Elabscience A3 Cell Line Manual

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Elabscience®

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A3 Cell Line Catalog No.: EP-CL-0315

Origin and General Characteristics Cell Name

A3 Cell Line

Organism

Homo sapiens, Human

Tissue

Peripheral blood

Morphology

Lymphoblast

Growth Properties

Suspension

Descriptions

The Jurkat cells were treated with Fas Antibody and isolated by limiting dilution to obtain a cell line that had a low spontaneous rate of resistance to Fas-medicated apoptosis. The resulting wild-type A3 subclone is very sensitive to Fas-mediated apoptosis. Wild-type A3 cells were made neomycin resistant and treated with three cycles of exposure to the frameshifting mutagen ICR-191 to isolate clones harboring recessive mutations that were resistant to killing by Fas antibody. ICR-191 treated clones were serially diluted in 96-well plates in the presence of Fas Antibody for 3 to 5 weeks. Two of these ICR-191 treated clones have been deposited at the ATCC. They are I 9. 2, a clone with a mutation in the cysteine protease caspase-8/FLICE and I 2. 1, a clone with a mutation in the adaptor FADD.

Biosafety Level

1

Culture Conditions and Handling Complete Growth Medium

RPMI-1640 +10% FBS +1% P/S

Subculturing

Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 to 2×10^5 viable cells/ml. Maintain cell density between 1×10^5 and 1×10^6 viable cells/ml.

Subcultivation Ratio

1:2-1:4

Medium Renewal

Every 2 to 3 days

Cryopreservation

Freeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase

Culture Conditions

Atmosphere: Air, 95%; CO2, 5% Temperature: 37℃

Recommendations for handling of cryopreserved cells 1.

The cell is packaged by dry ice. When receiving the cell, please make sure that the vial is still frozen. If there is cell thawing in the tube, please take photo before experiment or storage.

2.

If immediate culturing is not intended, the cryovial(s) must be stored in liquid nitrogen (-196°C) or at least at -80°C after arrival. If immediate culturing is intended, please follow these instructions:

3.

Quickly thaw by rapid agitation in a 37℃ water bath within 45-90 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath.

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