p65(RelA) Translocation Assay Kit (Part No. p65-Trans)

Manufacturer Fivephoton
Category Products / Kits
Type Kit
Specific against other
Amount 1 kit
ArtNr P65-Trans
Eclass 6.1 32161000
Eclass 9.0 32161090
Available
Highlights
Rapid, quantitative assay to measure translocation of p65 from cytoplasm to nucleus.
Generates a nuclear fraction with concentrated p65 protein.
This assay operates in most mammalian cell types.
Background
Following stimuli that elicit the NF-KB pathway, the IKB inhibitory subunit of the NF-kappa B complex is phosphorylated, ubiquitinated and degraded, exposing nuclear targeting signals in the transcription factor subunits of the NF-kappa B complex. The NF-kappa B transcription factor subunits (p65 and p50) with exposed nuclear import signals translocate into the nucleus and bind regulatory elements that modulate gene transcription. Since nuclear import is a key event in NF-kappa B activation, fractionation of cells into cytoplasmic and nuclear components and detection of translocated p65 in the nuclear fraction provides a quantitative assay for p65 translocation.

This p65(RelA) Translocation Assay Kit provides all of the necessary reagents to fractionate cells into cytoplasmic and nuclear fractions, as well as a selective and high affinity antibody to detect p65 in downstream Western blots.
p65(RelA)Translocation Assay Kit Contents
Cytoplasmic Fractionation Reagent (CER-1; 55 ml)
Nuclear Fractionation Reagent (NER-1; 3.5 ml)
DTT (sufficient to make a 1M solution with a volume of 64 microliters)
Rabbit Polyclonal IgG Antibody to p65. 100 microliters provided: Use at 1:400 dilution for Western blots
References
Fazal F, Bijli KM, Minhajuddin M, Rein T, Finkelstein JN, Rahman A. 2009. Essential role of cofilin-1 in regulating thrombin-induced RelA/p65 nuclear translocation and intercellular adhesion molecule 1 (ICAM-1) expression in endothelial cells. J Biol Chem 284: 21047-56.

Fazal F, Minhajuddin M, Bijli KM, McGrath JL, Rahman A. 2007. Evidence for actin cytoskeleton-dependent and -independent pathways for RelA/p65 nuclear translocation in endothelial cells. J Biol Chem 282: 3940-50.

Lim JW, Kim KH, Kim H. 2008. NF-kappaB p65 regulates nucleartranslocation of Ku70 via degradation of heat shock cognate protein 70 in pancreatic acinar AR42J cells. Int J Biochem Cell Biol 40: 2065-77.

Loveridge CJ, MacDonald AD, Thoms HC, Dunlop MG, Stark LA. 2008. The proapoptotic effects of sulindac, sulindac sulfone and indomethacin are mediated by nucleolar translocation of the RelA(p65) subunit of NF-kappaB. Oncogene 27: 2648-55.

Mattioli I, Sebald A, Bucher C, Charles RP, Nakano H, et al. 2004. Transientand selective NF-kappa B p65 serine 536 phosphorylation induced by T cell costimulation is mediated by I kappa B kinase beta and controls the kinetics of p65 nuclear import. J Immunol 172: 6336-44.
Amount: 1 kit
Available: In stock
Listprice: €470.08
Price: €470.08
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