NFkB, p65, phosphorylated (Ser529) (NF-kB, Nuclear Factor kappa B, RelA)

Nuclear factor-kB is a family of transcription factors that consists of homo- and heterodimers of\nNFkB1/p50 and RelA/p65 subunits, and controls a variety of cellular events including\ndevelopment and immune responses. All members share a conserved amino-terminus domain that includes dimerization, nuclear localization, and DNA binding regions, and a carboxy-terminal\ntransactivation domain. Serines 529 and 536 in the transactivation domain of RelA/p65 are\nphosphorylated in response to several stimuli including phorbol ester, IL-1b and TNF-a as\nmediated by IkB kinase and p38 MAPK. Serine 529 is located in a negatively charged region\n(amino acids 422-540) that is phosphorylated in response to phorbol myristate acetate plus calcium ionophore activation. Phosphorylation of serines 529 and 536 is critical for RelA/p65\ntranscriptional activity.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 \nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nJurkat cells treated with PMA and Ca2+ ionophore.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

N2302-04B

Size

10 Blots

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS (without Mg2+ and Ca2+), 1mg/ml BSA (IgG, protease free), pH 7.2, 0.05% sodium azide, 50% glycerol.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

References

Haller, D., et al. (2002) IKK beta and phosphatidylinositol 3-kinase/Akt participate in nonpathogenic\nGram-negative enteric bacteria-induced RelA phosphorylation and NF-kappa B activation in both primary and intestinal epithelial cell lines. J. Biol. Chem. 277(41):38168-38178. Hunter, R.B., et al. (2002) Activation of an alternative NF-kappaB pathway in skeletal muscle during disuse atrophy. FASEB J. 16(6):529-538. Martin, A.G., et al. (2001) Regulation of Nuclear factor kB transactivation. Implication of\nphosphatidylinositol 3-kinase and protein kinase C zeta in c-Rel activation by tumor necrosis factor\nalpha. J. Biol. Chem. 276 (19):15840-15849. Madrid, L.V., et al. (2001) Akt stimulates the transactivation potential of the RelA/p65 subunit of NF-kappa B through utilization of the Ikappa B kinase and activation of the mitogen-activated protein kinase p38. J. Biol. Chem. 276(22):18934-18940. Wang, D., et al. (2000) Tumor necrosis factor alpha-induced phosphorylation of RelA/p65 on Ser529 is controlled by casein kinase II. J. Biol. Chem. 275(42):32592-32597. Wang, D. and A.S. Baldwin, Jr. (1998) Activation of nuclear factor-kappaB-dependent transcription by tumor necrosis factor-alpha is mediated through phosphorylation of RelA/p65 on serine 529. J. Biol. Chem. 273(45):29411-29416.

Additional Info

Recognizes human NFkB when phosphorylated at Ser529. Species sequence homology: mouse NFkB (91%).