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

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