Cell Signaling Technology

Product Pathways - NF-kB Signaling

IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (Alexa Fluor® 488 Conjugate) #5743

Applications Reactivity Sensitivity Isotype
F H M R Mk B Pg GP Endogenous Mouse IgG1

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Pg=Pig  GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (Alexa Fluor® 488 Conjugate) detects endogenous levels of total IκBα protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a GST-IκBα fusion protein corresponding the amino terminus of human IκBα.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (Alexa Fluor® 488 Conjugate) (blue) compared to Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate) #4878 (red).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814.

Background

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).

  1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
  2. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
  3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
  4. Brown, K. et al. (1995) Science 267, 1485-8.
  5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  8. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.

Application References

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Alexa Fluor® is a registered trademark of Molecular Probes, Inc.Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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