Cell Signaling Technology

Product Pathways - NF-kB Signaling

Phospho-RelB (Ser552) Antibody #4999

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC F H M (R) (Mk) (B) (Dg) Endogenous 70 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Dg=Dog
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-RelB (Ser552) Antibody detects endogenous levels of RelB only when phosphorylated at Ser552.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser552 of mouse RelB. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from A20 and CTTL2 cells, untreated or treated with TPA #4174 (200nM, 30 minutes) alone or with λ phosphatase, using Phospho-RelB (Ser552) Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Raji cells, untreated (blue) or TPA-treated (green), using Phospho-RelB (Ser552) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Raji cells, serum-starved (left), treated with TPA (Phorbol-12-Myristate-13-Acetate) #9905 (center) or λ phosphatase-treated (right), using Phospho-RelB (Ser552) Antibody (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

Transcription factors of the nuclear factor κ B (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

RelB, which is generally activated by non-canonical signaling, forms heterodimers with either p50 or p52 NF-κB subunits to regulate transcription (12,13). RelB null mice are significantly impaired in inflammatory responses and hematopoietic differentiation (14,15). Phosphorlyation at Thr84 and Ser552 results in proteosomal degradation (16).

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.
  12. Ryseck, R.P. et al. (1992) Mol Cell Biol 12, 674-84.
  13. Bours, V. et al. (1994) Oncogene 9, 1699-702.
  14. Weih, F. et al. (1995) Cell 80, 331-40.
  15. Burkly, L. et al. (1995) Nature 373, 531-6.
  16. Marienfeld, R. et al. (2001) Oncogene 20, 8142-7.

Application References

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