Flow cytometric analysis of Jurkat cells, untreated (upper) or paclitaxel-treated (lower), using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb (Alexa Fluor® 488 Conjugate) versus propidium iodide (DNA content). The blue inserts represent propidium iodide staining alone, showing cells in mitotic arrest in the paclitaxel-treated sample.
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
posted July 2009
revised June 2017
Protocol Id: 407
Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb detects endogenous levels of Bcl-2 only when phosphorylated at Ser70. This antibody does not cross-react with nonphosphorylated Bcl-2 at endogenous levels or with other Bcl-2 family members.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser70 of human Bcl-2. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.
Bcl-2 exerts a survival function in response to a wide range of apoptotic stimuli through inhibition of mitochondrial cytochrome c release (1). It has been implicated in modulating mitochondrial calcium homeostasis and proton flux (2). Several phosphorylation sites have been identified within Bcl-2 including Thr56, Ser70, Thr74, and Ser87 (3). It has been suggested that these phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (6). Interleukin-3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced anti-apoptotic functions (7).
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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.