NF-kB p65, phosphorylated (Ser276) Blocking Peptide

Peptide is used to block Phospho-NFkB p65 (Ser276) Antibody reactivity in immunohistochemistry and Western blot protocols. \n\nTranscription factors of the nuclear factor kB (NF-kB)/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-kB1 (p105/p50) and NF-kB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. The p50 and p52 products form dimeric complexes with Rel proteins, which are then able to bind DNA and regulate transcription. In unstimulated cells, NF-kB is sequestered in the cytoplasm by its inhibitory proteins, the IkB's (3-5). NF-kB-activating agents can induce the phosphorylation of IkB's, targeting them for rapid degradation through an ubiquitin-proteasome pathway, releasing NF-kB to enter the nucleus, where it regulates gene expression (6-8). Processing of NF-kB2 p100 is regulated by NIK and IKK1 (IKKa), which triggers the phosphorylation and processing to p52, which can then undergo nuclear translocation (9-11).\n\nQuality Control: The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-NF-kB p65 (Ser276) Antibody by immunohistochemistry and Western blotting. \n\nDirections For Use: For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

N2304-10

Size

100ug

Applications

IHC

Form

Supplied as a liquid in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA and 5% glycerol.

Purity

Highly Purified

References

(1) Baeuerle, P.A. and Henkel, T. (1994) Annu. Rev. Im- munol. 12, 141-179. \n(2) Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20. \n(3) Haskill, S. et al. (1991) Cell 65, 1281-1289. \n(4) Thompson, J.E. et al. (1995) Cell 80, 573-582. \n(5) Whiteside, S.T. et al. (1997) EMBO J. 16, 1413-1426. \n(6) Traenckner, E.B. et al. (1995) EMBO J. 14, 2876-2883. \n(7) Scherer, D.C. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 11259-11263. \n(8) Chen, Z.J. et al. (1996) Cell 84, 853-862. \n(9) Senftleben, U. et al. (2001) Science 293, 1495-1499. \n(10) Coope, H.J. et al. (2002) EMBO J. 21, 5375-5385. \n(11) Xiao, G. et al. (2001) Mol. Cell 7, 401-409.