Flow cytometric analysis of HeLa cells, untreated (green) or treated with hTNF-α #8902 and Calyculin A #9902 (20 ng/ml and 100 nM, 15 min; blue), using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (Pacific Blue™ Conjugate) (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control (Pacific Blue™ Conjugate) #14411 (dashed line).
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This Cell Signaling Technology antibody is conjugated to Pacific Blue™ fluorescent dye and tested in-house for direct flow cytometry in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated antibody IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.
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
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (Pacific Blue™ Conjugate) detects endogenous levels of total IκBα protein.Species Reactivity:
Human, Mouse, Rat, Monkey, Bovine, Pig
Monoclonal antibody is produced by immunizing animals with a GST-IκBα fusion protein corresponding to the amino-terminus of human IκBα protein.
The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).
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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Pacific Blue is a trademark of Molecular Probes, Inc.