Cell Signaling Technology

Product Pathways - NF-kB Signaling

Phospho-IκBα (Ser32/36) (5A5) Mouse mAb (Sepharose Bead Conjugate) #4088

Applications Reactivity Sensitivity MW (kDa) Isotype
IP H M R Mk Dg (B) (Pg) Endogenous 40 Mouse IgG1

Applications Key:  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Dg=Dog  Pg=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

Phospho-IκBα (Ser32/36) (5A5) Mouse mAb (Sepharose Bead Conjugate) detects endogenous levels of IκBα only when phosphorylated at Ser32/36.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser32/36 of human IκBα.

IP

IP

Immunoprecipitation of NIH/3T3 cell lysates, untreated or TNF-α-treated, using Mouse IgG (Sepharose Bead Conjugate) #3420 (Lanes 1 and 2) and Immobilized Phospho-IκBα (Ser32/36) (5A5) Mouse mAb (Bead Conjugate) (Lanes 3 and 4). The blot was probed using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246.

Description

This Cell Signaling Technology antibody is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated sepharose beads. Phospho-IκBα (Ser32/36) (5A5) Mouse mAb (Sepharose Bead Conjugate) is useful for the immunoprecipitation of phosphorylated IκBα.

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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Companion Products

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

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