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MAP Kinase Pathways Research Tools

T

he mitogen-activated protein (MAP) kinases are a group of protein serine/threonine kinases that are activated in response to a variety of extracellular stimuli and mediate signal transduction from the cell surface to the nucleus.1,2 In combination with several other signaling pathways, they can differentially alter the phosphorylation status of transcription factors. The controlled regulation of MAP kinase cascades is involved in cell proliferation and differentiation, whereas an unregulated activation can result in oncogenesis.3-5

Three major types of MAP kinase cascades have been reported in mammalian cells that respond synergistically to different upstream signals. The most widely studied cascade is that of ERK1/ERK2 MAP kinases. A general activation scheme involves the activation of receptor tyrosine kinases by growth factors, such as EGF, which provides the binding site for the adapter protein Grb2 which in turn localizes Sos to the plasma membrane. Sos activates Ras by exchange of GTP for GDP. The RasGTP binds directly to a serine/threonine kinase, Raf, forming a transient membrane anchoring signal.6,8 Active Raf kinase phosphorylates a dual specificity kinase, MEK, on serine218 and serine222 and activates it.7 MEK can also be phosphorylated by Mos, a protein kinase expressed during meiotic maturation of oocytes and by MEKK1.8 A generally held belief is that MEK1 binds ERK and phosphorylates either a threonine or a tyrosine residue and then dissociates. The monophosphorylated ERK then rebinds to an active MEK1 for dual phosphorylation and complete activation.9,10 The activated MEK phosphorylates ERK1/ERK2 on threonine183 and tyrosine185 (at the TEY motif). The major targets of activated ERKs are pp90 ribosomal S6 kinase (Rsk) and the cytoplasmic phospholipase A2.11 ERK also translocates to the nucleus to phosphorylate the transcription factor Elk-1 (on serine383 and serine389). Recently another related kinase, ERK3, a nuclear protein kinase, has been cloned. It exhibits about 50% homology with ERK1/ERK2 within its catalytic domain.12 However, it does not phosphorylate any typical ERK substrates.

Usually only one highly active form of ERK1 or ERK2 (dual phosphorylated) exists in the cell, exhibiting over 1000-fold greater activity than the unphosphorylated form. Within the cell, at any time, one may find three less active forms of ERKs: one unphosphorylated enzyme, and two singly phosphorylated forms that contain phosphate either at the tyrosine or threonine residue.10 The JNK/SAPK (c-Jun kinase/stress activated protein kinase) cascade is activated following exposure to UV radiation, heat shock, or inflammatory cytokines.2,13 The activation of these MAP kinases is mediated by Rac and Cdc42, two small G-proteins.1,13 The activated Cdc42 binds to PAK65 protein kinase and activates it.14 The activated PAK65 can activate MEKK which in turn phosphorylates and activates SEK/ JNKK at serine219 and serine223.2 The active SEK/JNKK phosphorylates JNK/SAPK (at the TPY motif) that in turn binds to the Nterminal region of c-Jun and phosphorylates it at serine63 and serine73.16 The sites of activating phosphorylation are conserved between ERK and JNK, however, these sites are located within distinct dual specificity phosphorylation motifs (TPY for JNK and TEY for ERK).1,17 Molecular cloning studies show about 40 - 45% sequence homology between JNK/SAPK and the classical MAP kinases.16,18 The p38 kinase (reactivating kinase) is the newest member of the MAP kinase family.2 It is activated in response to inflammatory cytokines, endotoxins, and osmotic stress.5,18,19 It shares about 50% homology with the ERKs.5 The upstream steps in its activation are not well defined. However, downstream activation of p38 occurs following its phosphorylation (at the TGY motif) by MKK3, a dual specificity kinase. Following its activation, p38 translocates to the nucleus and phosphoryates ATF-2.20 Another known target of p38 is MAPKAPK2 that is involved in the phosphorylation and activation of heat-shock proteins.21


Although different MAP kinase cascades show a high degree of specificity and functional separation, some degree of cross-talk is observed between these pathways. For example, JNKK, an activator of JNK/SAPK, is reported to activate p38,22 whereas MKK3 activates only p38 and not JNK/SAPK. MEKK1, which stimulates SEK/JNKK1 in the JNK/SAPK cascade, has only a trivial effect on p38 activation. In the upstream signaling, Sos stimulates only the ERK pathways without affecting either JNK or p38 cascade. Another important observation is that in mammalian cells treated with mitogenic agents, ERKs are significantly activated whereas JNK/SAPK are not affected. Conversely, cells exposed to stress activate only the JNK/SAPK pathway without altering the activity of ERKs. At the transcription level, ATF-2 is phosphorylated and activated by all three MAP kinases, whereas c-Jun and Elk-1 are phosphorylated only by ERKs and JNK/SAPK, yet all these pathways result in transcriptional activity that is unique for a particular external stress.11 References: 1. Davis, R.J. 1994. Trends Biochem. Sci. 19, 470. 2. Su, B., and Karin, M. 1996. Curr. Opin. Immunol. 8, 402. 3. Sivaraman, V.S., et al. 1997. J. Clin. Invest. 99, 1478. 4. Das, R., and Vonderhaar, B.K. 1996. Breast Cancer Res.Treat. 40, 141. 5. Tong, L., et al. 1997. Nature Struct. Biol. 4, 311. 6. Winston, L.A., and Hunter, T. 1996. Curr. Biol., 6, 668. 7. Avruch, J., et al. 1994. Trends Biochem. Sci. 19, 279. 8. Sagata, N. 1997. BioEssays 19, 13. 9. Burack, W.R., and Sturgill, T.W. 1997. Biochemistry 36, 5929. 10. Cobb, M.H., et al. 1996. Adv. Pharmacol. 36, 49. 11. Waskiewicz, A.J., and Cooper, J.A. 1995. Curr. Opin. Cell Biol. 7, 798.

12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22.

Cheng, M., et al. 1996. J. Biol. Chem. 271, 8951. Hibi, M., et al. 1993. Genes Dev. 7, 2135. Martin, G.A., et al. 1995. EMBO J. 14, 1970. Kyriakis, J.M., et al. 1994. Nature 369, 156. Moriguchi, T., et al. 1996. Adv. Pharmacol. 36, 121. Derijard, B., et al. 1994. Cell 76, 1025. Han, J., et al. 1994. Science 265, 808. Rouse, J., et al. 1994. Cell 78, 1027. Raingeand, J., et al. 1995. J. Biol. Chem. 270, 7420. Larochelle, S., and Suter, B. 1995. Gene 163, 209. Lin, A., et al. 1995. Science 268, 286.

Activators Product

Cat. No.

Description

Size

Anisomycin, Streptomyces griseolus

176880

Strongly activates stress-activated protein kinases (JNK/SAPK) and p38 MAP kinase in mammalian cells. Synergizes with growth factors to induce c-Fos and c-Jun by acting as a potent signaling agonist. An inhibitor of protein synthesis at the translation step. Also known to induce apoptosis in the human monoblastoid cell line, U937.

10 mg

Aurintricarboxylic Acid

189400

Inhibits apoptotic cell death in various cell types induced by a variety of factors. A potent inhibitor of DNA topoisomerase II. Stimulates the tyrosine phosphorylation of MAP kinases, Shc proteins, PI 3-kinase, and PLC-g. Also a potent inhibitor of angiogenesis that potentiates the activity of angiostatic steroids. Inhibits both major calpain isoforms (IC50 = 22 mM and 10 mM for m-calpain and m-calpain respectively).

100 mg

Palytoxin, Palythoa toxica

506438

A non-protein toxin from sea anemones that is cytotoxic at picomolar concentrations. Produces multiple effects on both excitable and non-excitable cells. Activates stressactivated protein kinases/c-Jun N-terminal kinases (SAPK/JNK) in Swiss 3T3 fibroblasts. Depolarizes excitable cells and increases the intracellular Ca2+ in cardiac and smooth muscle. Binds to Na+,K+ pump and inhibits Na+,K+-ATPase activity thereby allowing K+ efflux from cells. A cation ionophore and non-TPA tumor promoter.

D-erythro-Sphingosine, N-Acetyl-

110145

Biologically active, cell-permeable, non-physiological ceramide analog. Inhibits cell growth and induces apoptosis in HL-60 cells. Induces intranucleosomal DNA fragmentation. Activates SAPK in HL-60 human promyelocytic cells. An activator of heterotrimeric PP2A. Stimulates a cytosolic Ser/Thr phosphatase in T9 cells at concentrations as low as 100 nM.

5 mg

D-erythro-Sphingosine, N-Hexanoyl-

376650

Biologically active, cell-permeable, non-physiological ceramide analog. Induces a dramatic arrest in the the cell cycle. Activates MAP kinase. Activates protein phosphatase 2A (PP2A) at 10 nM. Inhibits diacylglycerol accumulation and phospholipase D activation in fibroblasts. Induces apoptosis in Molt-4 leukemia cells and potentiates the activity of angiostatic steroids.

5 mg

D-erythro-Sphingosine, N-Octanoyl-

219540

A short chain cell-permeable analog of endogenous ceramide. Shown to induce apoptosis in a variety of cell lines and to activate MAP kinase in HL-60 cells.

5 mg

Sphingosylphosphorylcholine

567735

A deacetylated derivative of sphingomyelin known to accumulate in neuropathic 10 mg Niemann-Pick disease type A. Rapidly induces tyrosine phosphorylation of focal adhesion kinase (p125FAK) and paxillin and causes a rapid and transient activation of MAP kinase in Swiss 3T3 cells.A potent mitogen that increases intracellular free Ca2+ and free arachidonate through pathways that are only partially PKC-dependent.

Tumor Necrosis, Factor-a, Human, Recombinant, E. coli

654205

Activates a variety of immune defense mechanisms by interactions with polymorphonuclear leukocytes, T cells, antibody-producing B lymphocytes, fibroblasts, and hematopoietic bone marrow cells. Activity is not species-specific. Stimulates stress activated protein (SAP) kinase. Induces apoptosis in human blood and bone marrow neutrophils and in endothelial cells. Increases the iNOS levels in vascular smooth muscle cells. Involved in pathophysiological processes of several chronic and acute diseases.

25 mg 100 mg

10 mg


Inhibitors Product

Cat. No.

Description

Size

AG 126

658452

Blocks LPS-induced tyrosine phosphorylation of a p42 MAPK protein substrate. Inhibits lipoplysaccharide (LPS)-induced synthesis of TNF-Îą and nitric oxide in murine peritoneal macrophages.

5 mg

Apigenin

178278

Induces the reversion of transformed phenotypes of v-H-ras-transformed NIH 3T3 cells at low concentration (12.5 ÂľM), most likely by inhibiting MAP kinase activity. Inhibits the proliferation of malignant tumor cells by G2/M arrest and induces morphological differentiation.

5 mg

Hsp25 Kinase Inhibitor

385880

A 13-residue peptide that acts as a potent and selective inhibitor of mammalian heat-shock 1 mg protein (Hsp25) kinase [also called MAPKAP kinase-2]. Inhibition is competitive with respect to the substrate peptide (Ki = 8.1 mM) and non-competitive with respect to ATP (Ki = 134 mM).

Hypericin

400075

Inhibits protein kinase C (IC50 = 3.3 mM). Also inhibits casein kinase II (IC50 = 6 nM), MAP kinase (IC50 = 4 nM), and the protein tyrosine kinase activities of the insulin receptor (IC50 = 20 - 29 nM) and the EGF receptor (IC50 = 35 nM). Reported to inhibit PI-3 kinase activity (IC50 = 180 nM).

1 mg

5-Iodotubercidin

407900

A potent and competitive inhibitor of the MAP kinase ERK2 (Ki = 530 nM). Also inhibits adenosine kinase (Ki = 30 nM), casein kinase I and PKA, and the insulin receptor kinase (IC50 values range from 0.4 - 28 mM).

1 mg

MAP Kinase Antisense Oligonucleotide

454858

A 17-mer phosphorothioate oligodeoxynucleotide directed against a sequence that is identical in the p42 and p44 MAP kinases (ERK2 and ERK1). Causes a concentrationdependent decrease in the steady state expression of p42 and p44 MAP kinase in 3T3 L1 fibroblasts and adipocytes.

25 nmol

MAP Kinase Antisense Oligonucleotide, Control

454859

A 17-mer phosphorothioate oligodeoxynucleotide with the same nucleotide composition as the MAP kinase antisense oligonucleotide (Cat. No. 454858) but in a scrambled sequence. Does not affect MAP kinase expression or activity.

25 nmol

MAP Kinase Inhibitor Set

444180

Contains 5 mg of the MEK inhibitor PD98059 (Cat. No. 513000), 1 mg each of the MAP kinase inhibitors SB 202190 (Cat. No. 559388) and SB 203580 (Cat. No. 559389), and 1 mg of the negative control, SB 202474 (Cat. No. 559387). Supplied with an informational insert.

1 set

MEK Inhibitor Set

453710

Contains 5 mg of PD 98059 (Cat. No. 513000), 1 mg of U0126 (Cat. No. 662005) and 1 mg of the negative control U0124 (Cat. No. 662006). Supplied with an informational insert.

1 set

Olomoucine

495620

A purine derivative that acts as a potent and selective inhibitor of p34cdc2/cyclin B (IC50 = 7 mM) and related kinases including p33cdk2/cyclin A (IC50 = 7 mM), p33cdk2/cyclin E (IC50 = 7 mM), p33cdk5/p35 (IC50 = 3 mM), and p44 MAP kinase (IC50 = 25 mM). Acts by competing for the ATP binding domain of the kinase. Exhibits reduced sensitivity towards related kinases, p34cdk4/cyclin D (IC50 >1 mM) and p40cdk6/cyclin D3 (IC50 > 250 mM).

1 mg 5 mg

PD 98059

513000

Selective and cell-permeable inhibitor of MEK that acts by inhibiting the activation of MAP kinase and subsequent phosphorylation of MAP kinase substrates. Pretreatment of PC-12 cells with PD 98059 completely blocks the 4-fold increase in MAP kinase activity produced by nerve growth factor (NGF; IC50 = 2 mM). Does not affect the NGF-dependent tyrosine phosphorylation of the pp140trk receptor or its substrate Shc and does not block NGFdependent activation of PI 3-kinase.

5 mg

PD 169316

513030

A potent, cell-permeable, and selective p38 MAP kinase inhibitor (IC50 = 89 nM). Blocks apoptosis induced by trophic factor withdrawal in non-neuronal and neuronal cell lines.

1 mg

p38 MAP Kinase Inhibitor

506126

A potent p38 kinase inhibitor (IC50 = 35 nM).

500 mg

PD 184352

513045

A potent and selective MEK inhibitor. Directly inhibits MEK1 (IC50 = 17 nM) but does not affect MAP kinase (MAPK) activity. Does not affect other kinases such as MKK3, protein kinase C (PKC), cyclin-dependent kinase 2A, phosphorylase kinase, EGFR kinase, PDGF receptor kinase, or c-Src, at high concentrations (10 mM). Displays potent anti-tumor properties in vivo.

500 mg

SB 202190

559388

A potent and cell-permeable inhibitor of p38 MAP kinase. Inhibits p38 phosphorylation of 1 mg myelin basic protein (MBP) with no effect on the activity of the ERK or JNK MAP kinase subgroups. Also inhibits the kinase activity of p38b (IC50 = 350 nM) and p38 phosphorylation of activating transcription factor 2 (ATF-2; IC50 = 280 nM).

SB 202190, Hydrochloride

559393

A water-soluble form of the potent p38 MAP kinase inhibitor SB 202190 (Cat. No. 559388).

1 mg

SB 202474

559387

A negative control for MAP kinase inhibition studies.

1 mg


Inhibitors, cont. Product

Cat. No.

Description

Size

SB 203580

559389

A highly specific and cell-permeable inhibitor of p38 MAP kinase (IC50 = 600 nM). Suppresses the activation of MAPKAP kinase-2 and prevent the phosphorylation of heat shock protein 27 (Hsp27) produced in response to IL-1, cellular stresses, and bacterial endotoxin in vivo. Does not inhibit the MAP kinase homologs JNK or p42 MAP kinase.

1 mg

SB 203580, Hydrochloride

559395

A water-soluble form of the MAP kinase inhibitor SB 203580 (Cat. No. 559389).

1 mg

SB 203580, Iodo-

559400

A highly specific and potent inhibitor of p38 MAP kinase. Exhibits similar inhibitory potency as its parent compound SB 203580 (Cat. No. 559389). Useful for the identification of inhibitor binding sites in p38 MAP kinase.

1 mg

SB 220025

559396

A potent and specific inhibitor of human p38 mitogen-activated protein (MAP) kinase (IC50 = 60 nM). Exhibits over 2000-fold greater selectivity for p38 MAPK over ERK (p42/ p44 MAP kinase), 500-fold over PKA, 50-fold over PKC, and >1000-fold over EGFR. Also acts as a potent inhibitor of angiogenesis and as an inhibitor of TNF-a production.

500 mg

SC68376

565625

Potently and selectively inhibits p38 MAP kinase activity in a dose-dependent manner (IC50 = 2 - 5 mM).

1 mg

SKF-86002

567305

A bicyclic imidazole cytokine-suppressive, anti-inflammatory drug (CSAID) that inhibits osmotic stress and UV-induced apoptosis by blocking p38 MAP kinase activation as well as lipopolysaccharide (LPS)-stimulated IL-1 and TNF-a production (IC50 = 1 mM). Inhibits both cyclooxygenases and 5-lipoxygenase, and inhibits osmotic stress.

5 mg

U0124

662006

A useful negative control for MEK inhibitors U0125 (Cat. No. 662008) and U0126 (Cat. No. 662005). Does not inhibit MEK activity even at concentrations of 100 mM.

1 mg

U0125

662008

A specific inhibitor of MEK (MEK1 and MEK2). Although structurally similar to U0126 (Cat. No. 662005), it is about 10-fold less potent.

1 mg

U0126

662005

A potent and specific inhibitor of MEK1 (IC50 = 72 nM) and MEK2 (IC50 = 58 nM). The inhibition is noncompetitive with respect to MEK substrates, ATP, and ERK. Acts as an immunosuppressant by effectively blocking IL-2 synthesis and T cell proliferation without affecting the long-term outcomes of either T cell activation or tolerance.

1 mg

ZM 336372

692000

Inhibits SAPK2a/p38a (IC50 = 2 mM) and SAPK2b/p38b2 (IC50 = 2 mM). A potent inhibitor of c-Raf (IC50 = 70 nM).

1 mg

Substrates Product

Cat. No.

JNK Substrate, c-Jun (1-79)

420108

Description

Size

GST fusion protein of c-Jun (1 - 79) activation domain, expressed in E. coli. Highly specific substrate for c-Jun N-terminal kinase/stress activated protein kinase (JNK/SAPK). Is not phosphorylated by p42 MAP kinase or p38. Suitable for qualitative and quantitative assays of JNK activity.

100 mg

c-Jun (1-79)-GST, 420109 Human, Recombinant, E. coli, Agarose Conjugate

Purified c-Jun in a 50% agarose slurry. Agarose-bound form of c-Jun (1-79)-GST (Cat. No. 420103) useful for affinity purification of SAPK from cellular extracts. The phosphotransferase activity of bound SAPK then can be measured following the addition of g-32P-ATP.

100 mg

MAP Kinase Substrate, Tyrosine Hydroxylase 24-33

454860

Sequence features residues 24 - 33 of tyrosine hydroxylase, which corresponds to the consensus site for MAP kinase phosphorylation and includes the Ser31 phosphorylation site. Supplied as a trifluoroacetate salt.

1 mg

Myelin Basic Protein Peptide Substrate

475920

Substrate for p44mapk (ERK1) and p42mapk (ERK2) kinases. Corresponds to amino acid residues 94-102 of the myelin basic protein, which contains the consensus sequence Pro-X-(Ser/Thr)-Pro that is recognized by MAP Kinases. Phosphorylated by kinases on threonine97. Supplied as a trifluoroacetate salt.

1 mg

PHAS-I, Rat, Recombinant

516675

A 117-amino acid protein substrate which has been shown to be an excellent substrate for MAP kinase, p38 kinase, PKC, and JNK. Phosphorylation of PHAS-I increases in response to insulin but not to cAMP stimulation of adipocytes. An excellent substrate for MAP kinase both in vivo and in vitro.

250 mg


Antibodies Product Anti-ATF-1, N-Terminal (Sheep) Anti-ATF-2, Human (Rabbit) Anti-ERK1 (7-20), Rat (Rabbit)

Applications

Cat. No.

Size

IB, IP

114780

200 mg

IB, IP

189274

100 ml

ELISA, IB

442670

100 mg

442671

100 mg

442686

100 ml

ERK1 (7-20) Blocking Peptide, Rat Anti-ERK5, N-Terminal (1-14), Human (Rabbit)

ELISA, IB

ERK5, N-Terminal (1-14) Blocking Peptide, Human

442687

100 mg

Anti-MAP Kinase, Human (Rabbit)

IB, IC, IP

442704

200 ml

Anti-MAP Kinase, Phospho-Specific, Human (Rabbit)

IB, IC, IP

442705

200 ml

Anti-MAP Kinase-Activated Protein Kinase 2, C-Terminal (344-360), Human (Rabbit)

IB, IC, IP

442690

100 mg

442691

100 mg

442675

100 mg

442676

100 mg

442700

100 mg

MAP Kinase-Activated Protein Kinase 2, C-Terminal (344-360) Blocking Peptide, Human Anti-MAP Kinase ERK1/ERK2 (333-367), Rat (Rabbit)

ELISA, IB,IC,IP

MAP Kinase ERK1/ERK2 (333-367) Blocking Peptide, Rat (Rabbit) Anti-MAP Kinase ERK2 (Mouse) Anti-MAP Kinase ERK2, N-Terminal (6-17), Rat (Rabbit)

IB, IP IB

MAP Kinase ERK2, N-Terminal (6-17) Blocking Peptide, Rat Anti-MAP Kinase ERK3 (303-325), Rat (Rabbit)

IB

MAP Kinase ERK3 (303-325) Blocking Peptide, Rat Anti-MEK1, C-Terminal (359-377), Human (Rabbit)

IB, IH, IP

MEK1, C-Terminal (359-377) Blocking Peptide, Human Anti-MEK1, N-Terminal (1-12), Rat (Rabbit)

IB, IH, IP

MEK1, N-Terminal (1-12) Blocking Peptide, Rat

442680

100 mg

442681

100 mg

442683

100 mg

442684

100 mg

444946

100 mg

444948

100 mg

444942

100 mg

444943

100 mg

Anti-MEK1/MEK2, Human (Rabbit)

IB

444944

100 ml

Anti-MEK1/MEK2, Phospho-Specific, Human (Rabbit)

IB

444945

100 ml

Anti-MEK5 (59-74), Human (Rabbit)

ELISA, IB, IP

MEK5 (59-74) Blocking Peptide, Human Anti-MEK56 (320-334), Human (Rabbit)

IB, IP

Anti-MKK3, Human (Rabbit) Anti-cMos (149-177), Mouse (Rabbit)

444958

100 mg 100 mg

ELISA, IB, IP

453701

100 mg

IB

475884

100 ml

ELISA, IB

cMos (149-177) Blocking Peptide, Mouse Anti-p38/HOG1, C-Terminal (341-360), Human (Rabbit)

IB, IP

p38/HOG1, C-Terminal (341-360) Blocking Peptide, Human Anti-p38 MAP Kinase (Rabbit) Anti-p38 MAP Kinase, Phospho-Specific, Human (Rabbit) Anti-PHAS-I (Ab-1) (Rabbit)

100 mg 100 mg

444960

MEK56 (320-334) Blocking Peptide, Human Anti-MEKK1, N-Terminal, Rat (Rabbit)

444954 444956

475931

100 mg

475933

100 mg

506115

100 mg

506117

100 mg

IB

506123

200 ml

IB, IC, IP

506124

20 tests

IB, IP

PC256

100 mg

Anti-Raf1 (Ab-2) (Rabbit)

IB

PC91

100 mg

Anti-Raf1 (253-269), Human (Rabbit)

IB

Raf1 (253-269) Blocking Peptide, Human Anti-A-Raf Protein Kinase, C-Terminal (584-597), Human (Rabbit)

ELISA, IB, IP

A-Raf Protein Kinase, C-Terminal (584-597) Blocking Peptide, Human Anti-B-Raf Protein Kinase, C-Terminal (734-747) (Rabbit)

ELISA, IB, IH

B-Raf Protein Kinase, C-Terminal (734-747) Blocking Peptide

543501

100 mg

543503

100 mg

553001

100 mg

553002

100 mg

553004

100 mg

553006

100 mg

Anti-SEK1, C-Terminal (378-395), Mouse (Rabbit)

ELISA, IB, IP

563103

100 mg

SEK1, C-Terminal (378-395) Blocking Peptide, Mouse

ELISA, IB, IP

563107

100 mg

Inhibitors

ELISA: enzyme-linked immunosorbent assay; IB: immunoblotting; IC: immunocytochemistry; IH: immunohistochemistry; IP: immunoprecipitation.


Enzymes Product

Cat. No.

Description

Size

c-Jun N-Terminal Kinase, Rat, Recombinant

420105

Purified recombinant b-form of rat skeletal muscle c-Jun N-terminal kinase (JNK) expressed in E. coli. Involved in the cellular responses to stress signals such as UV irradiation, heat shock, and protein synthesis inhibitors by phosphorylating and enhancing the transcriptional activity of c-Jun. Other possible substrates of JNK include ATF2 and p53. Useful for the identification and characterization of JNK substrates and upstream activators of JNK. Capable of autophosphorylation.

20 mg

MAP Kinase, Mouse, Recombinant, E. coli

454850

Acts at the end of a protein kinase cascade by linking signals from cell surface receptor tyrosine kinases to cytoplasmic and nuclear events. This protein was co-expressed with MEK and is phosphorylated and active.

2000 U

MAP Kinase, Activated, Rat, Recombinant, E. coli

454855

Highly active form of MAP kinase produced by phosphorylating non-activated MAP kinase (Cat. No. 454852) in vitro with a constitutively active MEK mutant. Suitable for labeling MAP kinase substrates. Also useful for inhibitor screening and microinjection. Features a polyhistidine tag to facilitate removal of MAP kinase from the reaction mixture.

MAP Kinase, Non-Activated, Rat, Recombinant, E. coli

454852

Suitable to assay activity of MAP kinase activators such as MEK in cell lysates or in chromatography fractions. Features a polyhistidine tag to facilitate removal of MAP from kinase the reaction mixture.

MAP Kinase ERK1GST•Tag™, Human, Recombinant, Agarose Conjugate

442673

Purified human recombinant ERK1 in a 50% agarose slurry. A full length MAP kinase expressed as GST-fusion protein linked to glutathione-agarose. ERK1 can be released by incubating with bovine thrombin (Cat. No. 604980; 20 units/ml of slurry) or by eluting ERK1-GST with 10 mM glutathione. Useful as substrate for MEK1 and MEK2.

100 mg

[Ala71]-MAP Kinase ERK1, Inactive, Human, Recombinant, Agarose Conjugate

442674

Purified [Ala71]-ERK1 in a 50% agarose slurry. Mutation of Lys71 in ERK1 to an alanine results in a catalytically inactive form of the MAP kinase that can neither autophosphorylate nor phosphorylate other proteins. Suitable for use as a substrate for the detection and quantitation of MEK1 and MEK2.

100 mg

MEK1-GST•Tag™, Mouse, Agarose Conjugate

444952

Purified MEK1-GST in a 50% agarose slurry. This product will phosphorylate ERK1 and ERK2, and also serves as a substrate for MEKK (MAP kinase kinase kinase) and Raf1. Full-length soluble GST-MEK1 can be released by incubation of the agarose beads in 10 mM glutathione.

100 mg

MEK1 (K97A)GST•Tag™, Mouse, Agarose Conjugate

444941

Mouse MEK1 (K97A)-GST in a 50% agarose slurry. Mutation of Lys97 to Ala results in a catalytically inactive form of MEK that cannot phosphorylate ERK1 or ERK2. It is primarily used as a substrate for the detection and quantitation of MAP kinase kinase kinase (MEKK). Thus far, only MEK1 and MEK2 have been shown to serve as efficient substrates for Raf1.

100 mg

p38, GST•Tag™, Mouse, Recombinant

506122

Shares the highest homology (52% identity) with the yeast high osmolarity glycerol (HOG1) protein that mediates yeast osmosensing signaling. Recombinant p38 can phosphorylate ATF2 and PHAS-I (Cat. No. 516675) but not c-Jun (1-79) (Cat. No. 420108). Reported to phosphorylate PLA2 in thrombin-stimulated platelets. Also plays an important role in regulating inflammatory responses and apoptosis.

50 mg

SAPKb-GST•Tag™, Rat, Recombinant, E. coli, Agarose Conjugate

559310

A 50% agarose slurry. Cloned out of a rat brain cDNA library into the pGEX-KG GST fusion vector, expressed in E. coli, and linked to glutathione agarose beads. Primary utility of this product is the detection and quantitation of c-Jun. Full-length, soluble SAPKb can be released by incubation of the agarose beads with thrombin or the fusion protein can be released by incubation with 10 mM glutathione.

100 mg

SAPKb (K55R)-GST•Tag™, 559312 Rat, Recombinant, E. coli, Agarose Conjugate

A 50% agarose slurry. Cloned out of rat brain cDNA library into the pGEX-KG GST fusion vector, K55R mutated, expressed in E. coli, and linked to glutathione agarose beads. The mutation of Lys55 inactivates the SAPKb used for the detection and quantitation of c-Jun. Full-length, soluble SAPKb(K55R) can be released by incubation of the agarose beads with thrombin or the fusion protein can be released by incubation with 10 mM glutathione.

100 mg

SEK1 (K129R)-GST•Tag™, 563101 Mouse, Recombinant, E. coli, Agarose Conjugate

A 50% agarose slurry. Suitable for use as a substitute for MEKK and as a Western blot standard for Anti-SEK1 (Cat. No. 563103).

100 mg

5 mg

20 mg

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