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Immunohistology

Where Better Science Begins


®

HistoMark HistoMark Biotin Streptavidin Kits HistoMark Biotin Streptavidin Kits provide rapid, precise localization of antigens in cells and tissues. KPL offers two kits for use with a variety of primary antibodies: ♦ Biotin Streptavidin-HRP Systems ♦ Biotin Streptavidin-AP Systems KPL provides reagents optimized for sensitivity and convenience. They facilitate double or triple labeling experiments, as detection can be performed simultaneously with primary antibodies of different animal species. Contents of HistoMark Biotin Streptavidin Kits Reagents are supplied ready-to-use in convenient 50 mL dropper bottles with sufficient quantity to process 500 slides. Kits are stored at 2 - 8° C.

Each kit contains: Biotinylated Antibody With a long chain spacer arm that minimizes steric interaction with streptavidin and maximize the streptavidin-biotin binding reaction. Enzyme-labeled Streptavidin Designed for optimal binding to biotinylated antibody. Serum Block Minimizes nonspecific binding of the secondary antibody. KPL also offers HistoMark Staining Systems optimized for sensitivity and convenience. A variety of colors are available for both HRP and AP enzymes. They are ideal for multiple labeling experiments.

Images above: Left:Streptavidin Phosphatase and HistoMark RED demonstrate glial fibrillary acidic protein (GFAP) in brain. Middle: Streptavidin phosphatase and HistoMark BLUE demonstrates prostate-specific antigen in prostate. Right: Streptavidin Peroxidase and HistoMark ORANGE demonstrate epithelial membrane antigen in liver.

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HistoMark® Avidin-Biotin Complex (ABC) vs. Labeled Avidin Biotin (LAB)

ABC

Ordering Information Catalog No.

Description

Size

Biotin-Streptavidin - Phosphatase Systems 71-00-39 Mouse Primary Antibody 71-00-40 Rabbit Primary Antibody 71-00-41 Rat Primary Antibody

500 tests 500 tests 500 tests

Biotin-Streptavidin - Peroxidase Systems 71-00-18 Mouse Primary Antibody 71-00-19 Rabbit Primary Antibody 71-00-20 Rat Primary Antibody 71-00-26 Goat Primary Antibody

500 tests 500 tests 500 tests 500 tests

Components Available Separately 71-00-29 Goat Anti-Mouse IgG (H+L) 71-00-30 Goat Anti-Rabbit IgG (H+L), HSA 71-00-31 Goat Anti-Rat IgG (H+L), MSA 71-00-37 Rabbit Anti-Goat IgG (H+L) 71-0038 HRP-Streptavidin 71-00-45 AP-Streptavidin 71-00-27 Normal Goat Serum 71-00-28 Normal Rabbit Serum

50 mL 50 mL 50 mL 50 mL 50 mL 50 mL 50 mL 50 mL

HSA = Human Serum Adsorbed MSA = Mouse Serum Adsorbed

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

LAB

The technique utilizing unlabeled primary antibody, biotinylated link secondary reagent and pre-formed avidin-biotinylated horseradish peroxidase complex is known as the ABC method. (1) It has been reported that immunohisto-logic reactions using peroxidase coupled to streptavidin display greater sensitivity than ABC procedures. (2,3) In this method, the LAB method, the enzyme is covalently linked directly to the streptavidin molecule can be labeled with multiple enzymes. The resulting complex is relatively small, enabling many streptavidins to bind to the biotinylated secondary antibody. The biotin is also more accessible due to the use of a long carbon spacer arm which reduces potential steric hindrance. (1) Hsu, S-M, Raine, L. and Fanger, H. (1981) Am J Clin Pathol 75:734. (2) Shi, Z-R, Itkowitz, S.H. and Kim, Y.S. (1988) J Histochem Cytochem 36:317. (3) Elias, J.M., Margiotta, M. and Gaborc, D. (1989) Am J Clin Pathol 92:62

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

ISO 9001:2000 Registered

HistoMark is a registered trademark of KPL, Inc. © 2003 KPL, Inc. All rights reserved. ML-263-06 SEE MORE with KPL!


HistoMark® TrueBlue

Epithelial membrane antigen in kidney.

HistoMark® BLACK

T lymphocytes in tonsil.

StableDAB®

HistoMark® ORANGE

DAB Substrate

Smooth muscle actin in skin tissue.

Cytokeratin antigen in rat skin.

Retinal pigment epithelial cells, Class III β-tubulin.

HistoMark® BLUE

HistoMark® RED

Retinal glial cells, glial fibrillary acidic protein.

Prostate-specific antigen in prostate.

HistoMark® X-Gal

Keratin in skin.

HistoMark

®

HistoMark® Substrate and Staining Systems HistoMark® Staining Systems provide rapid, precise localization of antigens in cells and tissues. Systems are available for the detection of peroxidase, phosphatase and β-galactosidase-labeled

When used with KPL’s HistoMark Biotin Streptavidin Kits, they offer brilliant, intense colors and superior performance. ♦

Stable, liquid systems eliminate waste

reagents:

Convenient, easy-to-follow protocols

HistoMark Substrate

Α variety of contrasting colors for multiple staining

conjugates. Each kit contains the following

Stable liquid concentrates provide a variety of colors for multiple staining applications. Substrate Buffer Solution Maximizes color development Counterstain Provides sharp contrast and enhances nuclear detail.

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HistoMark Substrate and Staining Systems KPL’s staining systems offer a variety of contrasting colors for high resolution multiple labeling applications. KPL’s TrueBlue Substrate is highly recommended for in situ hybridization; a brilliant blue stain with superior resolution.

HistoMark® KPL’s HistoMark Biotin Streptavidin Kits provide

Cytomegalovirus-infected cells detected with a biotinylated CMV probe and the DNADetector Chromogenic in situ Hybridization Kit using TrueBlue.

StableDAB demonstrates cytokeratin 19 in breast tissue. TrueBlue demonstrates smooth muscle actin in breast tissue.

high signal-to-noise ratios when used with HistoMark Staining Systems.

KPL’s HistoMark Substrates and staining systems can be utilized to obtain brilliant

HistoMark BLUE demonstrates retinal pigment epithelial cells, Class III βtubulin. HistoMark RED demonstrates keratin.

double-staining results. To read more about immunohistochemistry double staining and for a guide to

Ordering Information Catalog No.

Description

“HistoMark Double Staining Size

“Technical Info” section at

For use with phosphatase 55-69-00 HistoMark RED 55-70-00 HistoMark BLUE

1000 slides 1000 slides

For use with peroxidase 54-74-00 HistoMark ORANGE 54-75-00 HistoMark BLACK 54-78-00 HistoMark TrueBlue 54-10-00 DAB Reagent Set 54-11-00 StableDAB Peroxidase Substrate

1000 slides 1000 slides 500 slides 500 slides 500 slides

For use with β-galactosidase 54-13-00 HistoMark X-Gal Substrate Set

200 slides

Counterstains 71-01-01 71-02-01 71-00-11 71-00-06 71-00-05

50 mL 10 mL 50 mL 50 mL 50 mL

Orcein Eosin Contrast GREEN Contrast BLUE Contrast RED

Procedures” visit the

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

www.kpl.com.

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

ISO 9001:2000 Registered

ML-264-05

StableDAB and HistoMark are registered trademarks of KPL, Inc. © 2003 KPL, Inc. All rights reserved.

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Cy3 and Cy5 Conjugates Image above: Confocal fluorescence micrograph of HeLa cells stained with monoclonal antibody against mitochondria enzyme and Cy3conjugated anti-mouse antibody (red); rabbit polyclonal antibody to histones in DNA and Cy5-conjugated anti-rabbit antibody (blue).

KPL CyTM Dye Conjugates- Improve your assay and brighten your day! Cy Dyes offer intense fluorescence when coupled

photostability. Cy Dye conjugates are used in both

with KPL’s affinity purified antibodies and strepta-

visual and image analysis fluorescent microscopy

vidin. The sensitivity and reproducibility of KPL

and in situ hybridization. Cy3 conjugates are ideally

antibodies combined with the brightness of Cy3 and

used in visual color applications, whereas Cy5 con-

Cy5 dyes produce an exceptional set of conjugates

jugates emit in the far red spectrum and are not easi-

ideal for multiple labeling experiments. Dye/protein

ly visualized.

ratios have been established to ensure optimal fluorescence with minimal background. They present maximum excitation/emission spectra at 550/570 nm (Cy3) and 650/670 nm (Cy5).

Cy3

Cy5

Benefits of KPL Cy Dye Conjugates High performance conjugates – optimized dyeprotein ratios ensure high signal-to-noise. Intense fluorescence – offers greater sensitivity than TRITC conjugates.

Ex Em

Narrow emission spectra – enables sensitive, multi-color analysis.

Fluorescence

Excellent photostability – more photostable than TRITC conjugates. Consistent performance – minimal lot-to-lot variation reduces need for assay optimization. Buffer stability – after rehydration, conjugates are stable at pH 4-9. Wavelength (nm)

Cy dyes are excellent alternatives to most other fluorrescein dyes as they are brighter and offer greater

Instrument compatibility – excitation and emission spectra correspond with standard filter sets and laser settings. SEE MORE with KPL!


Excellent Performance As demonstrated below KPL Cy3- and Cy5-labeled conjugates produce brighter fluorescence than those of other suppliers.

Cy 3 and Cy5 Conjugates

Cytomegalovirus-infected cells detected with a biotinylated CMV probe and the DNADetectorTM Fluorescent in situ Hybridization Kit using Cy3-Strept-avidin and DAPI.

Microplates were coated with serially diluted mouse IgG at the indicated concentrations. Conjugates at a concentration of 0.01 mg/mL were applied and incubated for 30 minutes. Fluorescence was measured with a Perkin Elmer VICTOR 3 Multilabel Plate Reader.

Ordering Information Description Anti-Mouse IgG (γ), HSA F(ab’)2 Anti-Mouse IgG (γ), HSA Anti-Mouse IgG (H+L), HSA F(ab’)2 Anti-Mouse IgG (H+L), HSA Anti-Mouse IgG (H+L), RbSA, HSA Anti-Mouse IgM (µ), HSA Anti-Mouse IgG+IgM (H+L), HSA Anti-Rabbit IgG (H+L) F(ab’)2 Anti-Rabbit IgG (H+L), HSA Anti-Rabbit IgG (H+L), HSA Anti-Rat IgG (H+L) F(ab’)2 Anti-Human IgG (H+L) Anti-Human IgG (γ) F(ab’)2 Anti-Human IgG (γ) Anti-Human IgM (µ) F(ab’)2 Anti-Human IgM (µ) Anti-Guinea Pig IgG (H+L) Anti-Chicken IgG (H+L) Anti-Horse IgG (H+L) Anti-Swine IgG (H+L) Anti-Dog IgG (H+L) Anti-Sheep IgG (H+L) Anti-Goat IgG (H+L) Streptavidin

Cy3 072-01-18-02 202-01-18-02 072-01-18-06 202-01-18-06 072-01-18-18 072-01-18-03 072-01-18-09 072-01-15-06 202-01-15-16 072-01-15-16 072-01-16-06 202-01-10-06 072-01-10-02 202-01-10-02 072-01-10-03 202-01-10-03 072-01-17-06 072-01-24-06 072-01-21-06 072-01-14-06 072-01-19-06 072-01-23-06 072-01-13-06 072-01-30-00

Cy5 072-02-18-02 202-02-18-02 072-02-18-06 202-02-18-06 072-02-18-18 072-02-18-03 072-02-18-09 072-02-15-06 202-02-15-16 072-02-15-16 072-02-16-06 202-02-10-06 072-02-10-02 202-02-10-02 072-02-10-03 202-02-10-03 072-02-17-06 072-02-24-06 072-02-21-06 072-02-14-06 072-02-19-06 072-02-23-06 072-02-13-06 072-02-30-00

Signal Detection Cy3 is excited maximally at 550 nm and fluoresces maximally at 570 nm. It is excited to about 50% of maximum with an argon laser (514 nm or 528 nm lines), or to about 75% of maximum with a helium/neon laser (543 nm line) or mercury lamp (546 nm line). Cy5 is excited maximally at 650 nm and fluoresces maximally at 670 nm. It is excited to about 98% of maximum with a krypton/argon laser (647 nm line) or to about 63% of maximum with a helium/neon laser (633 nm line). Cy5 produces minimal autofluorescence of biological specimens in this region of the spectrum. A confocal microscope equipped with the appropriate laser for excitation and a far-red detector enable double labeling with Cy3 and Cy5.

Gaithersburg, MD 20878

HSA=human serum adsorbed RbSA=rabbit serum adsorbed Cy Dye antibody conjugates are made in goat except anti-goat and anti-sheep antibodies made in rabbit. Supplied in 1 mg lyophilized form.

Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com ISO 9001:2000 Registered

To order or for more information on KPL’s line of unlabeled and conjugated affinity purified antibodies, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. ML368-01 For research use only. Cy Dye is a trademark of GE Healthcare. ©2009 KPL, Inc. All rights reserved.

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DyLightTM Fluorescent Conjugates KPL’s DyLightTM Conjugates - A Brilliant Choice! KPL’s DyLightTM conjugates offer a brilliant choice in a variety of multicolor detection applications, including fluorescence microscopy, flow cytometry, Western blotting, ELISA and array platforms. Our affinity purified antibodies combined with a series of outstanding DyLight dyes provide superior performance over conventional CyDyeTM fluors, fluorescein and rhodamine, with performance comparable to that of Alexa Fluor® dyes (Figure 1). Enjoy these advantages when you switch to KPL’s DyLight Conjugates:

KPL offers eight DyLight dyes, including 405, 488, 549, 594, 633, 649, 680 and 800 with well-differentiated excitation and emission spectra. Our extensive line of over 170 DyLight conjugates is available across a range of animal species immunoglobulin, including human, mouse, rabbit, rat, other species and streptavidin. See back cover to find out what sets KPL’s antibodies apart. Comparison of DyLight 649 and Alexa 647 Conjugates in a Performance Assay

• High sensitivity, bright signal enable conjugate conservation

• More photostable than Cy, FITC; comparable to Alexa High signal-to-noise ratios; low background Wide selection of specific antibodies Consistent, lot-to-lot performance Narrow emission spectra enable specific, multicolor analysis • Compatible with a variety of buffers.

• • • •

Antigen Concentration (ng/mL)

Figure 1. KPL DyLight 649 conjugates demonstrate comparable fluorescense intensity and photostability to Alexa 647 conjugates in a FLISA.

Image above: HMVEC-L primary endothelial cells. F-actin detected with DY554-Phalloidin (rendered green). Microtubules detected with anti-tubulin antibody and DyLight 649 conjugated goat anti-mouse IgG (red). Nuclei detected with DAPI (blue)

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DyLightTM Fluorescent Conjugates DyLight Conjugated Affinity Purified Antibodies and Streptavidin KPL DyLight conjugates provide a choice of outstanding fluorescent conjugates with absorption spectra ranging from 405 nm to 800 nm. Their emission profiles correspond to those of other commonly used fluorophores such as Alexa, FITC and the Cy dyes. Narrow, defined emission spectra enable the use of multiple fluorescence labeling and simultaneous identification of several target molecules in the same sample. Light output is comparable to IRDye and Alexa and more intense than Cy Dyes, FITC or TRITC. DyLight dyes demonstrate resistance to photobleaching, resulting in excellent photostability as well as high solubility in aqueous solutions across a range of pH values.

Emission

DyLight Dye

Ex/Em(nm) Replaces

Blue

DyLight 405

400/420

Alexa 405 and Cascade Blue

DyLight 488

493/518

Alexa 488, Fluorescein and FITC

DyLight 549

550/568

Alexa 546, Alexa 555, Cy3, TRITC

Green Yellow Orange

DyLight 594

593/618

Alexa 594, Texas Red

Red

DyLight 633

638/658

Alexa 633

Red

DyLight 649

646/674

Alexa 647, Cy5

Near IR

DyLight 680

682/715

Alexa 680, Cy5.5

Infrared

DyLight 800

770/794

IRDye 800

nm = nanometers IR = infrared

Ordering Information Description Anti-Mouse IgG (γ), HSA Anti-Mouse IgG (H+L), HSA Anti-Mouse IgG (H+L), HSA, 0.1 mg Anti-Mouse IgG (H+L), RbSA, HSA Anti-Mouse IgM (µ), HSA Anti-Mouse IgG+IgM (H+L), HSA F(ab’)2 Anti-Mouse IgG (γ), HSA F(ab’)2 Anti-Mouse IgG (H+L), HSA Anti-Human IgG (H+L) Anti-Human IgG (H+L), 0.1 mg Anti-Human IgG (γ) Anti-Human IgM (µ) F(ab’)2 Anti-Human IgG (γ) F(ab’)2 Anti-Human IgM (µ) F(ab’)2 Anti-Human IgG (H+L) Anti-Rabbit IgG (H+L) Anti-Rabbit IgG (H+L), 0.1 mg Anti-Rabbit IgG (H+L), HSA F(ab’)2 Anti-Rabbit IgG (H+L), HSA Anti-Rat IgG (H+L) Anti-Rat IgG (H+L), 0.1 mg Anti-Guinea Pig IgG (H+L) Anti-Chicken IgG (H+L) Anti-Dog IgG (H+L) Anti-Goat IgG (H+L) Anti-Horse IgG (H+L) Rabbit Anti-Sheep IgG (H+L) Anti-Swine IgG (H+L) Streptavidin Streptavidin, 0.1 mg

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DyLight 405* DyLight 488 DyLight 549

072-08-18-06 042-08-18-06

202-08-18-06 072-08-10-06 042-08-10-06

202-08-10-06 072-08-15-06 042-08-15-06 202-08-15-16 072-08-16-06

072-08-24-06 072-08-13-06

072-08-30-00 042-08-30-00

*Coming soon!

072-03-18-02 072-03-18-06 042-03-18-06 072-03-18-18 072-03-18-03 072-03-18-09 202-03-18-02 202-03-18-06 072-03-10-06 042-03-10-06 072-03-10-02 072-03-10-03 202-03-10-02 202-03-10-03 202-03-10-06 072-03-15-06 042-03-15-06 072-03-15-16 202-03-15-16 072-03-16-06 042-03-16-06 072-03-17-06 072-03-24-06 072-03-19-06 072-03-13-06 072-03-21-06 072-03-23-06 072-03-14-06 072-03-30-00 042-03-30-00

DyLight 594*

DyLight 633* DyLight 649

072-04-18-02 072-04-18-06 042-04-18-06 072-04-18-18 072-04-18-03 072-04-18-09 202-04-18-02 202-04-18-06 072-04-10-06 042-04-10-06 072-04-10-02 072-04-10-03 202-04-10-02 202-04-10-03 202-04-10-06 072-04-15-06 042-04-15-06 072-04-15-16 202-04-15-16 072-04-16-06

072-05-18-02 072-09-18-06 072-10-18-06 042-09-18-06 042-10-18-06 072-05-18-18 072-05-18-03 072-05-18-09 202-05-18-02 202-09-18-06 202-10-18-06 072-09-10-06 072-10-10-06 042-09-10-06 042-10-10-06 072-05-10-02 072-05-10-03 202-05-10-02 202-05-10-03 202-09-10-06 202-10-10-06 072-09-15-06 072-10-15-06 042-09-15-06 042-10-15-06 072-05-15-16 202-09-15-16 202-10-15-16 072-09-16-06 072-10-16-06

072-04-17-06 072-04-24-06 072-04-19-06 072-04-13-06 072-04-21-06 072-04-23-06 072-04-14-06 072-04-30-00 042-04-30-00

072-05-17-06 072-09-24-06 072-10-24-06 072-05-19-06 072-09-13-06 072-10-13-06 072-05-21-06 072-05-23-06 072-05-14-06 072-09-30-00 072-10-30-00 042-09-30-00 042-10-30-00

DyLight 680

DyLight 800

072-05-18-06 042-05-18-06 072-06-18-18

072-06-18-06 042-06-18-06 072-07-18-18

072-07-18-06 042-07-18-06

202-05-18-06 072-05-10-06 042-05-10-06

202-06-18-06 072-06-10-06 042-06-10-06

202-07-18-06 072-07-10-06 042-07-10-06

202-05-10-06 072-05-15-06 042-05-15-06 072-06-15-16 202-05-15-16 072-05-16-06

202-06-10-06 072-06-15-06 042-06-15-06 072-07-15-16 202-06-15-16 072-06-16-06

202-07-10-06 072-07-15-06 042-07-15-06

072-06-17-06 072-05-24-06 072-06-19-06 072-05-13-06 072-06-21-06 072-06-23-06 072-06-14-06 072-05-30-00 042-05-30-00

072-07-17-06 072-06-24-06 072-07-19-06 072-06-13-06 072-07-21-06 072-07-23-06 072-07-14-06 072-06-30-00 042-06-30-00

202-07-15-16 072-07-16-06

072-07-24-06 072-07-13-06

072-07-30-00 042-07-30-00


Near Infrared and Infrared Fluorophores DyLight 680 and 800 Conjugates Offer Sensitive Multicolor Imaging in Western Blotting

DyLight Dyes Emission Spectra

DyLight 680 and 800 dyes emit in the near infrared and infrared ranges of the light spectrum respectively and are ideal for multicolor protein detection in Western blotting. Unlike fluorescent conjugates that emit in the visible range, DyLight 680 and 800 conjugates provide a unique set of advantages: • Secondary antibodies with defined specificity and

sensitivity • Brighter signal than visible fluorescence • Virtually no background autofluorescence from membranes or most biological specimens Effective Alternative to Chemiluminescence • Broader dynamic range than chemiluminescence • Quantitation accuracy superior to traditional

methods • Easy to assay multiple proteins simultaneously on one Western blot • Cost-effective - eliminates need for chemilumin-

escence substrates, film and darkroom • Clean results - no bleeding from consecutive lanes

Near Infrared Fluorescent Imaging with Secondary Antibodies labeled with DyLight 680 and 800

Figure 2. DyLight 680-anti mouse IgG and DyLight 800 anti-rabbit IgG secondary antibody conjugates provide low background and high signal in two-color Western blot detection of tubulin and TNFα. Membrane was imaged with the LI-COR Odyssey Infrared Imaging System. HSA = human serum adsorbed RbSA = rabbit serum adsorbed DyLight antibody conjugates are made in goat except anti-goat and anti-sheep antibodies are made in rabbit. Supplied in 1.0 mg lyophilized form except select 0.1 mg sizes.

1.800.638.3167

www.kpl.com


Immunofluorescence using DyLight Conjugates See bright fluorescence and low background with KPL DyLight conjugates in immunohistology applications. The winning combination of DyLight dyes and KPL purified antibodies enables multicolor labeling of two or more targets with similar intensity and photostability to Alexa dyes without the limitations of fluorescein and CyDyes.

Mouse primary cortical neurons MCx WCS stained with DyLight 549 (red). Synaptophysin stained with DyLight 488 (green).

HMVEC-L primary endothelial cells F-actin detected with DY554-Phalloidin (rendered green). Microtubules stained with DyLight 649 (red). Nuclei detected with DAPI (blue).

Rat hippocampal neurons WCS stained with; DyLight 488 (green). MAP2 stained with DyLight 549 (red).

A549 cells Cytokeratin stained with DyLight 680 (pseudocolored white). Lamin A stained with DyLight 549 (pseudocolored red).

DyLightTM Fluorescent Conjugates KPL AntibodiesWhat Sets Them Apart In 1979, KPL pioneered the production of large-scale affinity purification and was the first company to commercialize affinity purified secondary antibodies. Rigorous standards throughout the antibody production process make our antibodies standout in the marketplace. Many manufacturers cut corners by beginning with inferior serum and extracting the useful antibody towards the end of their process. KPL spends considerable effort developing and purifying its own immunogen formulation to generate the antiserum, because pure immunogen results in a more potent and specific antibody prior to any purification steps. Further, ISO 9001:2008-certified quality procedures are carried out at more than six different stages of the antibody production cycle, and material that does not meet our high standards for potency and cross-reactivity is rejected. Our late stage purification process has been continually refined since 1979 and relies on a one of a kind custom column resin which is uniquely suited to our antibody manufacturing process. Finally, our process of pooling antiserum from multiple animals tempers natural serum variability, minimizing variances from animal-to-animal. The result is more standardized large-scale antibody lots with increased consistency. KPL’s experience, innovative processes, and attention to detail result in highperformance antibodies that are unique.

NIH 3T3 cells F-actin detected with DY554-Phalloidin (rendered green). Microtubules stained with DyLight 649 (red). Nuclei detected with DAPI (blue) Gaithersburg, MD Phone: 800.638.3167/301.948.7755 Fax: 301.948.0169 www.kpl.com

To order or for more information on KPL’s line of unlabeled and conjugated affinity purified antibodies, contact us at 800.638.3167/301.948.7755,

ISO 9001:2008 Registered

fax 301.948.0169 or visit www.kpl.com. ML356-02 For research use only. DyLight is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries. Cy and Cy Dye are trademarks of GE Healthcare. Alexa Fluor is a registered trademark of Invitrogen. 2009 KPL, Inc. All rights reserved.

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ELISpot ELISpot Protein DetectorTM ELISpot Kits Protein Detector ELISpot kits are designed to provide complete, flexible systems for conducting ELISpot assays with any matched pair of antibodies. For the beginner or the more experienced researcher, these kits are ideal due to their comprehensive reagents and optimized protocol. Convenience Each kit contains all reagents for ELISpot detection except for matched antibody pairs (unlabeled coating and biotinylated antibody). Stable, ready-to-use, one-component substrates and uncoated Multiscreen速 PVDF Plates are included. Other kit components are supplied as concentrates; simply dilute and use. Stability All kit reagents are stable for a minimum of one year, providing reliable signal development and consistently low background.

Visualization Visualizing immunospots has never been easier with the exceptional localization and resolution provided by kit substrates. For sharp localization and fine resolution of bright blue immunospots, choose the HRP kit with TrueBlueTM substrate. For excellent visibility from a dark purple precipitate, choose the AP kit with BCIP/NBT substrate. Flexibility Universal nature of reagents allows kits to be used with any matched pair of antibodies. Since labeled streptavidin is compatible with any biotinylated antibody, the researcher simply provides the antibody pair for detection of different cytokines or other secreted proteins. Additional flexibility is offered through two kit formats. Choose between the HRP kit with TrueBlue or the AP kit with BCIP/NBT.

Image above. Mouse spleen cells secreting IFN-gamma were detected in ELISpot with either BCIP/NBT (top rows) or TrueBlue Substrate (bottom rows).

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ELISpot

Protein Detector HRP and AP ELISpot Kits Protein DetectorTM ELISpot Kits offer pretested reagents and a convenient, optimized protocol for ELISpot protein detection via either horseradish peroxidase (HRP) or alkaline phosphatase (AP) detection systems. With all necessary reagents except the paired antibody set (unlabeled capture antibody and biotinylated detection antibody), this kit allows for minimal time and effort in developing a sensitive and reproducible ELISpot assay. Provided conjugates can detect any biotinylated antibody making these reagents easily interchangeable with any matched antibody pair.

Related Products *TrueBlue Peroxidase Substrate Catalog No. 71-00-64 Size: 50 mL

*BCIP/NBT Substrate (1-Comp) Catalog No. 50-81-18 Size: 100 mL

Kit contents: • Chromogenic Substrate HRP Kit: TrueBlue AP Kit: BCIP/NBT • Labeled Streptavidin HRP Kit: Peroxidase labeled Streptavidin AP Kit: Phosphatase labeled Streptavidin • 5 Multiscreen PVDF Plates, 96-well • Coating Solution Concentrate • BSA Blocking/Diluent Solution Concentrate • Washing Solution Concentrate

HRP-labeled Streptavidin Catalog No. 474-3000 Size: 1.0 mL

AP-labeled Streptavidin Catalog No. 475-3000 Size: 1.0 mL

Comparison of Substrates for ELISpot Color Development

Substrate

Format

Enzyme

Sensitivity

TrueBlue

1-C

HRP

Blue

√√√√

BCIP/NBT

1-C

AP

Dark Purple

√√

AEC

2-C

HRP

Red

√√

C= Component

√=Least sensitive

√√√√=Most Sensitive

*BSA Diluent/Blocking Solution Concentrate Catalog No. 50-61-00 Size: 200 mL

Coating Solution Concentrate Catalog No. 50-84-00 Size: 50 mL

Wash Solution Concentrate Catalog No. 50-63-00 Size: 800 mL

* Other sizes available

Ordering Information Catalog#

Description

Size

40-00-05

Protein Detector HRP ELISpot Kit

5 plates

50-00-05

Protein Detector AP ELISpot Kit

5 plates

Each kit includes substrate (TrueBlue or BCIP/NBT), HRP or AP-labeled Streptavidin, 5 Multiscreen PVDF Plates, 96-well, Coating Solution Concentrate, BSA Blocking/Diluent Solution Concentrate, and Washing Solution Concentrate. Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

www.kpl.com

ISO 9001:2000 Registered

TrueBlue is a registered trademark and Protein Detector and Detector are trademarks of KPL, Inc. Multiscreen is a registered trademark of Millipore Corporation. © 2003 KPL, Inc. All rights reserved. ML290-05

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StableDAB® Liquid Diaminobenzidine for Immunohistochemistry and Blotting Diaminobenzidine (DAB) has long been a standard in peroxidase localization IHC applications. KPL’s StableDAB® offers excellent signal-to-noise in a liquid format that remains active and does not precipitate. It eliminates much of the handling required by traditional DAB. Revolutionary Stability Make StableDAB just once every two weeks! Mix 2 liquid components and use it for up to a full week when stored at room temperature, or up to 2 weeks if stored at 4° C.

Unbeatable Convenience Mix liquid chromogen with the liquid buffer and use for the next two weeks. Drastic Reduction in Exposure StableDAB minimizes handling. No thawing, weighing, dissolving, nor daily disposal. Use it longer, make it and dispose of it less often, reduce the risk to you and to the environment.

Superior Performance StableDAB forms a sharply localized brown deposit at the reaction site and is optimized for maximum signal and minimal background.

Image above: HRP-Streptavidin, StableDAB and Contrast Blue counterstain demonstrate smooth muscle actin in skin tissue.

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StableDAB® Superior Performance in Immunohistochemistry ™

StableDAB, when used with TrueBlue , offers convenient double staining with high resolution and contrast. KPL’s TrueBlue offers 100 times the sensitivity of DAB. High contrast and resolution of cellular details make this an excellent substrate for multiple staining procedures. Primary antibody can be diluted 10-50 times further than with DAB alone. TrueBlue is a convenient single component solution and poses no hazard to you or the environment. Both TrueBlue and StableDAB are alcohol and xylene insoluble. KPL offers a technical manual with double staining protocols using HistoMark® Immunohistochemical

StableDAB is packaged conveniently for staining individual or batches of slides, for manual and automated staining: ♦ 100 mL stabilized hydrogen peroxide substrate buffer at working dilution ♦ 2 mL DAB Concentrate in a dropper vial (sufficient to stain at least 500 slides).

Staining Systems.

Ordering Information Catalog No.

Description

Size

54-11-00

StableDAB

100 (mL)

HistoMark Immunohistochemical Staining Systems PEROXIDASE 54-10-00 DAB Reagent Set 50-78-02 TrueBlue 54-75-00 HistoMark RED 54-74-00 HistoMark ORANGE

500 slides 200 500 500

PHOSPHATASE 55-69-00 55-70-00

HistoMark RED HistoMark BLUE

500 500

β-GAL 54-13-00

HistoMark X-Gal Substrate Set

100

HistoMark Universal Streptavidin Kits Each kit contains serum block, biotinylated secondary antibody, and labeled streptavidin.

HistoMark Streptavidin Peroxidase Kits for Use with 71-00-18 Mouse Primary Antibody 71-00-19 Rabbit Primary Antibody 71-00-20 Rat Primary Antibody 71-00-26 Goat Primary Antibody

500 Slides 500 500 500

HistoMark Streptavidin Phosphatase Kits for Use with 71-00-39 Mouse Primary Antibody 71-00-40 Rabbit Primary Antibody 71-00-41 Rat Primary Antibody

500 500 500

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

ISO 9001:2000 Registered

StableDAB and HistoMark are registered trademarks and TrueBlue is a trademark of KPL, Inc. © 2003 KPL, Inc. All rights reserved. ML-205-09

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TrueBlue™ TrueBlueTM Peroxidase Substrate for Immunohistochemistry TrueBlue is a highly sensitive chromogenic substrate for visualization of horseradish peroxidase-labeled reporter reagents. It provides a brilliant blue specific stain, which contrasts sharply with red counterstains. TrueBlue Substrate is a buffered solution containing 3, 3’, 5, 5’-tetramethylbenzidine (TMB) and Hydrogen Peroxide (H2O2). TrueBlue offers the following benefits:

Economical TrueBlue permits extremely high dilutions of costly primary antibodies. You can perform many more assays with TrueBlue, saving you time and money. This stable, liquid system eliminates waste and reduces preparation time.

Ideal for Multiple Staining Highest Sensitivity TrueBlue is 50 to 100 times more sensitive than DAB. It provides an excellent means for detecting low abundant target antigens.

TrueBlue forms a dark blue reaction product that offers outstanding contrast with DAB and other substrates in sequential localization assays.

Safe–to–Use Highly Stable and Convenient TrueBlue is non-carcinogenic and safe-to-use, no special handling and disposal required.

TrueBlue is provided as a stable, singlecomponent working solution that is readyto-use. No reagent preparation required.

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TrueBlue™ Technical Tips TrueBlue Comparison to DAB

• Primary antibodies must be diluted 10 to 50 times further than with DAB, otherwise sample will overstain. Excess substrate may wash off the section, causing the appearance of understaining.

Methodology TrueBlue and DAB were compared using increasing dilutions of monoclonal antibody to latent cytomegalovirus (CMV) antigen on CMV-infected, acetone-fixed fibroblast cell monolayers. Monolayers were immunostained using identical protocols (monoclonal antibody and conjugate incubations of 30 minutes each, and substrate incubations of 10 minutes each).

• Color development in less than 10 minutes indicates that the primary antibody is too concentrated. • Blocking is essential because TrueBlue is extremely sensitive. Trace levels of peroxidase and nonspecific binding sites in the tissue must be neutralized to prevent background staining.

Conclusion

• Peroxidase block solution must be thoroughly removed by washing; it can affect antibody activity.

TrueBlue exhibited 100 times higher sensitivity than DAB. When using TrueBlue, dilute primary antibodies 10 to 50 times further for superior staining and lower background.

• KPL recommends non-aqueous mounting reagent. Fading will occur if aqueous media are used. • Stained sections must be completely dry or color may fade rapidly after mounting.

Ordering Information Catalog No.

Description

Size

50-78-02

TrueBlue Peroxidase Substrate

200 mL

71-00-64

TrueBlue Peroxidase Substrate

50 mL

71-00-67

TrueBlue Peroxidase Substrate

10 mL

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

www.kpl.com

ISO 9001:2000 Registered

ML-269-05

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Immunoassay Buffers Save preparation time with KPL’s new line of quality buffers! Lighten your work load with KPL’s new line of quality

registered quality management system. All buffers are con-

buffers. Offered as convenient liquid concentrates, they

ductivity controlled. KPL offers a variety of buffers designed

eliminate the need for time-consuming buffer preparation.

for use in general immunoassay applications such as Western

KPL buffers are manufactured and carefully controlled for

blotting, gel electrophoresis, ELISA and sample preparation.

quality and consistent performance with our ISO 9001:2008Description

1X Composition

Applications

10X Tris-Glycine Transfer Buffer

25 mM Tris, 192 mM glycine. pH 8.3

10X Tris-Glycine-SDS

25 mM Tris, 192 mM glycine, 0.1% SDS. pH 8.3

For preparing a standard Western blot transfer buffer (Towbin) and as a gel electrophoresis buffer for native Tris-glycine gels without SDS. . Running buffer for sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE) of proteins.

10X Phosphate-Buffered Saline (PBS)

10 mM Phosphate,150 mM NaCl. pH 7.4

Phosphate-Buffered Tween 20, (PBST)

10 mM Phosphate, 150 mM NaCl, 0.05% Tween 20. pH 7.4

10X Tris-Buffered Saline (TBS)

50 mM Tris, 150 mM NaCl. pH 7.6

10% Tween 20

10% Tween 20

Wash buffer for general immunoassay applications and sample preparation. Where applicable, Tween 20 is a nonionic detergent that reduces nonspecific binding and protein-protein interaction during wash steps.

Nonionic detergent additive for PBS and Western blotting wash solutions. Reduces nonspecific binding and proteinprotein interactions.

See reverse side.

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Immunoassay Buffers

KPL Quality Buffers (cont’d) Description

1X Composition

Applications

5% SDS

5% Sodium Dodecyl Sulfate

Detergent surfactant used in preparing proteins for sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE) and as an additive to transfer buffers in Western blotting. SDS increases the elution rate of proteins from the gel.

20X SSC

15 mM sodium citrate, 150 mM NaCl in DEPC water. pH 7.0

Used in a variety of molecular biology applications. Facilitates transfer of nucleic acids to membranes.

10X Dulbecco's PBS (D-PBS)

8.5 mM sodium phosphate, 1.5 mM potassium phosphate, 137 mM NaCl. pH 7.4

For sample preparation and as a wash buffer in general immunoassay, tissue, and cell culture applications. Not formulated with magnesium or calcium salts. Where applicable, Tween 20 is a non-ionic detergent that reduces nonspecific binding.

Other Immunoassay Support Reagents Available from KPL ELISA and Western Blotting Applications: • AP and HRP Conjugate Stabilizers

10X Dulbecco's with Tween 20. (D-PBST)

8.5 mM sodium phosphate, 1.5 mM potassium phosphate, 137 mM NaCl, 2.7 mM KCl. 0.05% Tween 20. pH 7.4

10X Tris-Buffered Saline with 0.5% Tween 20 (TBST)

50 mM Tris, 150 mM NaCl, 0.05% Tween 20. pH 7.6

10X Tris-Buffered Saline with 10% Tween 20 (TBST)

50 mM Tris, 150 mM NaCl, 1.0% Tween 20. pH 7.6

• 10% BSA Diluent/Blocking Solution Concentrate • Milk/Diluent Blocking Solution Concentrate • 20X Wash Solution Concentrate

As a general wash buffer in immunoassay applications. Tween 20 is a non-ionic detergent that reduces nonspecific binding. The 10% Tween 20 formulation provides a more stringent wash than standard TBST formulations.

Western Blotting Applications: • Detector Block • Biotin Wash Kit

Take the time and effort out of your buffer preparation. Choose from our line of popular buffers and enjoy the benefits - convenience, reliability and confidence - that KPL’s pretested, quality buffers bring to your research.

ELISA Applications: • Coating Solution Concentrate • Stop Solutions

Ordering Information Catalog#

Description

Size

51-10-01 51-10-02 51-11-01 51-11-02 51-13-01 51-13-02 51-14-01 51-14-02 51-17-01 51-17-02 51-12-01 51-12-02 51-20-01 50-86-05 51-15-01 51-15-02 51-16-01 51-16-02 51-18-01 51-18-02 51-19-01 51-19-02

10X Tris-Glycine Transfer Buffer

1L 5L 1L 5L 1L 5L 200 mL 1L 1L 5L 200 mL 1L 200 mL 1L 1L 5L 200 mL 1L 200 mL 1L 200 mL 1L

10X Tris-Glycine SDS Buffer 10X Phosphate-Buffered Saline (PBS) 10X Phosphate-Buffered Saline with Tween 20 (PBST) 10X Tris-Buffered Saline (TBS) 10% Tween 20 5% SDS 20X SSC 10X Dulbecco's PBS (D-PBS) 10X Dulbecco's PBS with Tween 20 (D-PBST) 10X Tris Buffered Saline with 0.5% Tween 20 (TBST) 10X Tris Buffered Saline with 10% Tween 20 (TBST)

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

ISO 9001:2008 Registered

To order or for more information on KPL’s full line of protein detection products, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. ©2009 KPL, Inc. All rights reserved. ML363-03

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Gold Conjugates Easy to use, highly sensitive systems for visualizing proteins Versatile and Convenient Probes

Silver Enhancer Kits

KPL's line of colloidal gold-labeled reagents include anti-human, anti-mouse, anti-rabbit, anti-biotin antibodies and streptavidin. These antibodies are affinity purified to minimize cross-reactivity. Anti-biotin and streptavidin conjugates provide a sensitive indication of biotin with low background.

For maximum signal, KPL's Silver Enhancer Kits amplify the gold conjugates signal 10 to 100 times. The resulting brown/black silver stain is permanent and offers sharp resolution and contrast with conventional counterstains.

Advantages of gold conjugates

Applications for Gold Conjugates 40 nm gold conjugates are ideal for use in these applications.

• Different sized gold particles (5 nm and 40 nm) allow for a variety of uses for these conjugates.

• Lateral flow assays

• Greater stability in liquid form than enzyme conjugates.

• Immunohistochemistry and in situ hybridization applications

• Eliminates the need for an enzyme blocking step. • 5 nm gold particles permit better penetration of tissue due to their small size. • Highly sensitive technique detects 60 picograms of protein in dot blotting.

• Light Microscopy • Blotting

5 nm gold conjugates are ideal for use in these applications. • Light Microscopy • Electron Microscopy

SEE MORE with KPL!


Gold Conjugates FREQUENTLY ASKED QUESTIONS

CMV infected cells, human anti-CMV serum with gold-labeled goat anti-human IgG (H+L), 5 nm with Microscopy Silver Enhancer Kit and Eosin.

Human normal lung tissue, anti-epithelial membrane antigen with gold-labeled goat anti-mouse IgG (H+L), 5 nm with Microscopy Silver Enhancer Kit and Contrast BLUE.

Ordering Information Catalog#

Description

Size

Gold Labeled Each system contains Gold Conjugate Dilution Buffer 57-10-06 Anti-Human IgG (H+L) System, 40 nm 58-10-06 Anti-Human IgG (H+L) System, 5 nm 57-18-06 Anti-Mouse IgG (H+L) System, 40 nm 58-18-06 Anti-Mouse IgG (H+L) System, 5 nm 57-15-06 Anti-Rabbit IgG (H+L) System, 40 nm 58-15-06 Anti-Rabbit IgG (H+L) System, 5 nm 57-40-06 Anti-Biotin System, 40 nm 58-40-06 Anti-Biotin System, 5 nm 57-30-06 Streptavidin System, 40 nm 58-30-06 Streptavidin System, 5 nm

What are the advantages of using gold conjugates versus enzyme-labeled conjugates? One advantage is the possibility of cross-reactivity with naturallyoccurring enzymes is non-existent, as compared to enzyme-labeled conjugates. Another advantage is a single-step detection versus enzyme conjugate and substrate.

3.0 mL 1.0 mL 3.0 mL 1.0 mL 3.0 mL 1.0 mL 3.0 mL 1.0 mL 3.0 mL 1.0 mL

Silver Enhancer Kits 55-22-01

55-22-02

Silver Enhancer Kit for Microscopy Kit includes: 1 mL of Gold Conjugate 100 mL of Gold Conjugate Dilution Buffer Solution A Solution B Silver Enhancer Kit for Membrane Applications Kit includes: 3 mL of Gold Conjugate 250 mL of Gold Conjugate Dilution Buffer Solution A Solution B

25 mL 25 mL

250 mL 250 mL

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

To order or for more information on KPL’s full line of protein detection products, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com.

ISO 9001:2000 Registered

Pictured on front and above: adult grasshopper. Š 2005 KPL, Inc. All rights reserved. ML248-06

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Peroxidase Conjugates KPL Peroxidase Conjugates: Time-tested, Sensitive and Reliable Since 1979 KPL has provided quality affinity

streptavidin. They are affinity purified and in

purified antibodies to researchers worldwide.

some cases further adsorbed to minimize cross-

Over the years we have refined our production

reactivity between animal species or shared

process to provide antibodies with high potency

reactivity with other immunoglobulin classes.

and consistent performance in immunoassays.

HRP-labeled F(ab’)2 fragment antibodies are

From the start KPL gives careful consideration

offered for assays requiring extremely low back-

to immunogen preparation, using a highly

ground and absence of F(c)-mediated binding.

purifed formulation to generate antiserum. KPL pools antiserum from multiple animals to

KPL reacts HRP of the highest quality with

reduce natural animal to animal serum variabili-

affinity purified antibodies and streptavidin

ty. During the purification process our ISO

using the periodate method of Nakane and

9001:2008-certified quality procedures impose

Kawaoi. Special features of HRP include:

rigorous standards for potency and cross-reac-

• faster catalytic rate than alkaline phosphatase • generates more product in shorter incubation

tivity. The result is standardized antibody lots with excellent reproducibility. Our extensive line of peroxidase (HRP) conjugates is available across a range of animal species, including human, mouse, rabbit and rat antibodies, as well as other animal species and

times

• provides maximum sensitivity, low nonspecific binding

• ideal for ELISA, Western blotting and immunohistology applications. SEE MORE with KPL!


Peroxidase Conjugates Peroxidase Conjugates Ordering Information (Partial listing) Catalog#

Description

Size

KPL offers a range of sensitive substrates for use with HRP conjugates. They provide a choice of intense colors for ELISA, blotting and cell staining applications.

04-10-06

HRP-labeled Goat Anti-Human IgG (H+L)

0.1 mg

04-10-17

HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L), MSA

0.1 mg

04-10-20

HRP-labeled Goat Anti-Human IgG (Fc)

0.1 mg

074-1002

HRP-labeled Goat Anti-Human IgG (γ)

1.0 mg

074-1003

HRP-labeled Goat Anti-Human IgM (μ)

1.0 mg

074-1004

HRP-labeled Goat Anti-Human IgE (ε)

1.0 mg

074-1006

HRP-labeled Goat Anti-Human IgG (H+L)

1.0 mg

074-1007

HRP-labeled Goat Anti-Human IgA+IgG+IgM (H+L)

1.0 mg

14-10-01

HRP-labeled Goat Anti-Human IgA (α)

0.5 mg

214-1002

HRP-labeled F(ab’)2 Goat Anti-Human IgG (γ)

0.5 mg

214-1003

HRP-labeled F(ab’)2 Goat Anti-Human IgM (μ)

0.5 mg

214-1006

HRP-labeled F(ab’)2 Goat Anti-Human IgG (H+L)

0.5 mg

474-1002

HRP-labeled Goat Anti-Human IgG (γ), Liquid

1.0 mL

474-1003

HRP-labeled Goat Anti-Human IgM (μ), Liquid

1.0 mL

474-1006

HRP-labeled Goat Anti-Human IgG (H+L), Liquid

1.0 mL

4 CN Peroxidase Substrate

04-18-06

HRP-labeled Goat Anti-Mouse IgG (H+L), HSA

0.1 mg

04-18-15

HRP-labeled Goat Anti-Mouse IgG (H+L), RtSA, HSA

0.1 mg

TMB Membrane Peroxidase Substrate

04-18-18

HRP-labeled Goat Anti-Mouse IgG (H+L), RbSA, HSA

0.1 mg

074-1802

HRP-labeled Goat Anti-Mouse IgG (γ), HSA

1.0 mg

074-1803

HRP-labeled Goat Anti-Mouse IgM (μ), HSA

1.0 mg

074-1806

HRP-labeled Goat Anti-Mouse IgG (H+L), HSA

1.0 mg

074-18-061

HRP-labeled Goat Anti-Mouse IgG (H+L), XSA

1.0 mg

074-1807

HRP-labeled Goat Anti-Mouse IgA+IgG+IgM (H+L), HSA

1.0 mg

074-1809

HRP-labeled Goat Anti-Mouse IgG+IgM (H+L), HSA

1.0 mg

14-18-01

HRP-labeled Goat Anti-Mouse IgA (α), HSA

0.5 mg

214-1802

HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (γ), HSA

0.5 mg

214-1806

HRP-labeled F(ab’)2 Goat Anti-Mouse IgG (H+L), HSA

0.5 mg

474-1802

HRP-labeled Goat Anti-Mouse IgG (γ), HSA, Liquid

1.0 mL

474-1806

HRP-labeled Goat Anti-Mouse IgG (H+L), HSA, Liquid

1.0 mL

074-1506

HRP-labeled Goat Anti-Rabbit IgG (H+L)

1.0 mg

074-15-061

HRP-labeled Goat Anti-Rabbit IgG (H+L), XSA

1.0 mg

074-1516

HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA

1.0 mg

214-1516

HRP-labeled F(ab’)2 Goat Anti-Rabbit IgG (H+L), HSA

0.5 mg

474-1506

HRP-labeled Goat Anti-Rabbit IgG (H+L), Liquid

1.0 mL

474-1516

HRP-labeled Goat Anti-Rabbit IgG (H+L), HSA, Liquid

1.0 mL

14-30-00

HRP-labeled Streptavidin

0.5 mg

Phone: 800.638.3167

474-3000

HRP-labeled Streptavidin, Liquid, Molecular Grade

1.0 mL

Fax: 301.948.0169

Visit our website at www.kpl.com for a complete listing of HRP-labeled antibodies. To order or for more information on KPL’s protein research products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

ELISA •

ABTS® 1- and 2-Component Microwell Peroxidase Substrates

SureBlueTM TMB Peroxidase Substrate

SureBlue ReserveTM TMB Peroxidase Substrate

TMB Peroxidase Substrate

Blotting

LumiGLO®Chemiluminescent Substrate LumiGLO ReserveTM Chemiluminescent Substrate

Whichever substrate you choose, enjoy the benefits of excellent signalto-noise and reproducibility.

Gaithersburg, MD 20878

www.kpl.com

ISO 9001:2008 Registered

For research use only. ©2009 KPL, Inc. All rights reserved. ABTS is a registered trademark of Roche Biochemicals. ML371-01

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in situ Hybridization DNADectectorTM in situ Hybridization Kits DNADetector in situ Hybridization Kits provide simple sensitive detection of DNA in cells, tissues, and metaphase chromosomes. Available for both chromogenic and fluorescent methods, these convenient kits include everything you need for in situ hybridization and detection of DNA except for biotinylated probe. DNA Detector Chromogenic in situ Hybridization Kit:

Biotin-labeled primary antibody must be supplied by kit user. DNADetector Fluorescent in situ Hybridization Kit: • Cy3 conjugates provide 5 - 10 times greater fluorescence than fluorescein-labeled probes1.

• Brilliant blue stain gives excellent contrast to red counterstains.

• Cy3 conjugates are more photostable than FITC or TRITC. Bright fluorescence can still be seen 6 months after staining.

• TrueBlueTM substrate provides superior resolution; very fine precipitate vs. large clumps of color with BCIP/NBT.

• No new filters required. Cy3 conjugates are visualized using the standard filters for TRITC.

• Sensitivity equal to that of FISH with permanent results.

• Kit contains all reagents, including control slides to perform in situ hybridization. Biotin-labeled primary antibody must be supplied by kit user.

• Kit contains all reagents, including control slides to perform in situ hybridization.

Photo on right: Cytomegalovirus infected cells detected with a biotinylated CMV probe and the DNADectector Chromogenic in situ Hybridization Kit using HRP-Streptavidin, TrueBlue and Orcein. Photo on left: Cytomegalovirus infected cells detected with a biotinylated CMV probe and the DNADetector Fluorescent in situ Hybridization Kit using Cy3-Streptavidin and DAPI. SEE MORE with KPL! 1 Yurov, Y. B. et. al. Human Genetics, 97, 390-398 (1996)


in situ Hybridization DNADectectorTM in situ Hybridization Kits

Related Products Universal Block Catalog No. 71-00-61 Size: 100 mL Herring Sperm DNA Catalog No. 60-00-14 Size: 40 mg

Herpes simplex virus infected tissue detected with a biotinylated herpes probe and the DNADetector Chromogenic in situ Hybridization Kit using TrueBlue and Orcein.

Cytomegalovirus infected cells detected with a biotinylated CMV probe and the DNADetector Fluorescent in situ Hybridization Kit using Cy3Streptavidin and DAPI.

DNADetector Chromogenic in situ Hybridization Kit

DNADetector Fluorescent in situ Hybridization Kit

Contents: Hybridization Buffers 20X SSC HRP Streptavidin Conjugate Wash Solution Conjugate Diluent buffer TrueBlue Peroxidase Substrate Orcein and Eosin Counterstains Control Hybridization Cocktail Control Slide

Contents: Hybridization Buffers 20X SSC Cy3 Streptavidin Conjugate Wash Solution Conjugate Diluent buffer DAPI Counterstain Fluorescent Mounting Media Control Hybridization Cocktail Control Slide

Sufficient reagents are provided for the hybridization

Sufficient reagents are provided for the hybridization

and detection of 50 samples.

and detection of 50 samples.

Fluorescent Mounting Media Catalog No. 70-00-16 Size: 15 mL Orcein Catalog No. 71-01-01 Size: 50 mL Eosin Y Catalog No. 71-02-01 Size: 10 mL DAPI Counterstain Catalog No. 71-03-01 Size: 1.0 mg

Ordering Information Catalog#

Description

Size

60-03-00 Chromogenic in situ Hybridization Kit 60-05-00 Fluorescent in situ Hybridization Kit, Cy3/DAPI Detector Probe Labeling Kits

50 samples 50 samples

60-01-00 60-01-01 60-01-02

30 reactions 30 reactions 20 reactions

Random Primer Biotinylation Kit PCR Biotinylation Kit RNA in vitro Transcription Kit

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169

To order or for more information on KPL’s full line of protein and nucleic acid detection products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com.

www.kpl.com

ISO 9001:2000 Registered

DNADetector is a trademark of KPL, Inc. Cy is a trademark of Amersham International PLC. Š 2003 KPL, Inc. All rights reserved. SEE MORE with KPL! ML224-11 1203


SureLINKTM Fluorescein Labeling Kits Simple, fast, convenient fluorescein labeling kits KPL offers two new SureLINKTM Fluorescein Labeling Kits

FAM-X and FITC share the same excitation (495 nm) and

that enable scientists to label proteins with the fluo-

emission (520 nm) wavelengths. However, FAM-X forms

rophores FITC or FAM-X (FAM fluorophore with succin-

a stable bond with the amine group of the protein that is

imidyl ester and a seven atom aminohexanoyl spacer,

more resistant to hydrolysis, minimizing the release of dye

known as ‘X’):

during immunoassay procedures and storage. In addition,

SureLINKTM Fluorescein (FITC)

the tendency towards quenching observed with FITC conjugates is diminished with FAM-X due to the spacer separating the fluorophore from the conjugated biomolecule.

Convenient kits save time and reagents Optimized protocols enable labeling and purifying in just two hours with minimal hands-on time. Kits contain reacFluorescein isothiocyanate (FITC)

tion-size reagent vials with all the essential components.

SureLINK Fluorescein-X (FAM-X)

Comprehensive conjugation guide A thorough guide is provided with both kits, and includes protocols, frequently asked questions and a troubleshooting guide.

Fluorescein X (FAM-X): 6-(fluorescein-5-carboxamido) hexanoic acid, succinimidyl ester SEE MORE with KPL!


SureLINKTM Labeling Kits

FITC and FAM-X Performance Comparison

Figure 1: Microwell plates were coated with 40 Âľg/mL of mouse IgG serially diluted across the plate. FITC-labeled anti-mouse IgG and fluorescein-Xlabeled anti-mouse IgG were added at a concentration of 500 ng and the plate was measured.

KPL offers a series of SureLINK Labeling Kits that enable fast, reliable labeling of antibodies and proteins through primary amine groups, including: SureLINK AP Conjugation Kits Catalog #85-00-02 SureLINK HRP Conjugation Kits Catalog #84-00-02

The results demonstrate that FAM-X achieves higher

SureLINK Chromophoric Biotin Labeling Kits

photoluminescence intensity than standard FITC.

Catalog #86-00-01

Fast and Convenient SureLINK Conjugation Protocol 1. Add buffer to protein in reaction tube.

2. Dissolve SureLINK fluorophore in DMF.

3. Aliquot into reaction tube and mix.

4. Remove unconjugated fluorophore with spin filter.

Total time = 1.5 - 2 hours Hands-on time = 30 minutes

1 hour

Successful labeling is easy with optimized kits that deliver speed and convenience. All necessary reagents for labeling proteins are provided. Reaction-size vials make it possible to conjugate a range of protein sample sizes. The conjugates are ideal for use in a variety of applications, such as ELISA, Western blotting, protein microarrays and immunohistochemistry.

~30 minutes

Visit kpl.com to learn more about our line of protein labeling kits.

Ordering Information Catalog#

Description

Size

82-00-01

SureLINKTM Fluorescein (FITC) Labeling Kit

5 reactions

82-00-02

SureLINKTM Fluorescein-X (FAM-X) Labeling Kit

5 reactions

SureLINK Kit Components SureLINK Fluorescein (FITC) Labeling Kit: SureLINK FITC, 5 x 0.3 mg 5 Carbonate Bicarbonate Buffer Capsules SureLINK Fluorescein-X (FAM-X) Labeling Kit: SureLINK Fluorescein-X (FAM-X), 5 x 0.3 mg Borate Buffer Both kits: Anhydrous DMF, 5 Spin-Pure Filters, 10 Amber Reaction Tubes

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com ISO 9001:2000 Registered

To order or for more information, contact us at 800.638.3167 / 301.948.7755, fax 301.948.0169 or visit us at www.kpl.com. ML350-01 For research use only. Š2007 KPL, Inc. All rights reserved. SEE MORE with KPL!


ReserveAP Conjugates TM

TM

Spice up your assay with our red hot high potency ReserveAP Conjugates! KPL’s new ReserveAPTM Alkaline Phosphatase

line of AP conjugates in ELISA and outperform

(AP)-labeled antibody conjugates exhibit high

AP conjugates offered by other manufacturers.

potency and consistent performance in

Higher conjugate dilutability is also observed

immunoassays. These conjugates are the result

without loss of linearity, enabling precious

of advances in our conjugation technology and

antigen or primary antibody conservation.

offer higher signal than our current line of AP

Consistent Performance

conjugates while meeting the same standards for low background, stability and reproducibili-

Reproducible antibody conjugation and consis-

ty. They are intended for demanding

tent performance are verified according to our

immunoassays that require high detection sen-

ISO 9001:2000-registered quality assurance

sitivity, including ELISA, Western blotting and

system. Lot consistency studies in which three

immunohistology.

lots were studied by ELISA indicated minimal variability.

Higher Potency Excellent Value ReserveAP Conjugates are affinity purified and conjugated to the highest grade of alkaline

ReserveAP Conjugates provide high perform-

phosphatase. In our studies, they generate two-

ance at an economical price.

to-three fold higher values than our current SEE MORE with KPL!


AP Substrates Chemiluminescent ELISA: A constant amount of antigen (mouse IgG) was coated onto a microtiter plate. The plate was probed with varying concentrations of goat anti-mouse IgG (H+L) AP conjugate, including KPL’s standard AP conjugate and ReserveAP™ conjugate, followed by KPL’s PhosphaGLO™ Chemiluminescent AP Substrate.

KPL offers a range of sensitive substrates for the detection and quantification of phosphatase (AP) activity. They provide a choice of intense colors for ELISA and blotting applications.

ELISA •

Conclusion: The results indicate that ReserveAP™ conjugate offers superior performance to our standard AP conjugate with a ~3-fold lower amount of conjugate required for detection.

ReserveAPTM Conjugates Ordering Information

Catalog#

Description

Size

0751-1001

Goat Anti-Human IgA (α)

1.0 mg

0751-1002

Goat Anti-Human IgG (γ)

1.0 mg

0751-1003 0751-1004

Goat Anti-Human IgM (µ) Goat Anti-Human IgE (ε)

0751-1006

FirePhosTM AP Microwell Substrate BluePhos® AP Microwell Substrate pNPP Phosphatase Substrate

Blotting •

1.0 mg 1.0 mg

FirePhos AP Membrane Substrate

BCIP/NBT Phosphatase Substrate

Goat Anti-Human IgG (H+L)

1.0 mg

PhosphaGLO AP Substrate

0751-1007

Goat Anti-Human IgA+IgG+IgM (H+L)

1.0 mg

2151-1002

0.5 mg

4751-1002

F(ab’)2 Anti-Human IgG (γ) Goat Anti-Human IgG (γ) liquid

PhosphaGLOTM Reserve AP Substrate

4751-1003

Goat Anti-Human IgM (µ) liquid

1.0 mg

4751-1006

Goat Anti-Human IgG (H+L) liquid

1.0 mg

0751-1802

Goat Anti-Mouse IgG (γ) HSA

1.0 mg

0751-1803

Goat Anti-Mouse IgM (µ) HSA

1.0 mg

0751-1806

Goat Anti-Mouse IgG (H+L) HSA

1.0 mg

0751-1809

Goat Anti-Mouse IgG +IgM (H+L) HSA

1.0 mg

4751-1802

Goat Anti-Mouse IgG (γ) HSA, liquid

1.0 mg

4751-1806

Goat Anti-Mouse IgG (H+L) HSA, liquid

1.0 mg

1.0 mg

151-18-01

Goat Anti-Mouse IgA (α) HSA

0.5 mg

0751-1807

Goat Anti-Mouse IgA+IgG+IgM (H+L) HSA

1.0 mg

0751-1506

Goat Anti-Rabbit IgG (H+L)

1.0 mg

0751-1516

Goat Anti-Rabbit IgG (H+L) HSA

1.0 mg

4751-1506

Goat Anti-Rabbit IgG (H+L), liquid

1.0 mg

4751-1516

Goat Anti-Rabbit IgG (H+L) HSA, liquid

1.0 mg

Whichever substrate you choose, enjoy the benefits of excellent signalto-noise with higher sensitivity and lower background than that of other AP substrates. Visit www.kpl.com for more information.

Gaithersburg, MD 20878 Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com ISO 9001:2000 Registered

HSA=Human Serum Adsorbed

Visit our website at www.kpl.com for a complete listing of ReserveAP conjugates. To order or for more information on KPL’s protein research products, contact us at 800.638.3167 / 301.948.7755, FAX 301.948.0169 or visit us at www.kpl.com. ML349-04 For research use only. ©2007 KPL, Inc. All rights reserved.

SEE MORE with KPL!


SEE MORE with KPL! Get specific signal without background and SEE MORE!

Where Better Science Begins

KPL, Inc. Gaithersburg, MD Phone: 800.638.3167 Fax: 301.948.0169 www.kpl.com

ML329-01


Immunohistology Kits and Reagents