The NF-κB p65 antibody is a crucial tool for studying the NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling pathway, which regulates immune responses, inflammation, apoptosis, and cell proliferation. The p65 subunit, also known as RelA, is a key component of the canonical NF-κB pathway. It forms heterodimers with p50. and in resting cells, these complexes are sequestered in the cytoplasm by inhibitory proteins like IκB. Upon stimulation by cytokines, pathogens, or stress signals, IκB is phosphorylated and degraded, enabling p65/p50 translocation to the nucleus to activate target genes.
Antibodies targeting p65 are widely used to detect its expression, localization, and activation status in various experimental models. Applications include Western blotting, immunohistochemistry (IHC), immunofluorescence (IF), and chromatin immunoprecipitation (ChIP). Phospho-specific antibodies (e.g., against Ser536) distinguish activated p65. providing insights into pathway dynamics. These antibodies help elucidate NF-κB's role in diseases like cancer, autoimmune disorders, and chronic inflammation.
Researchers must consider species reactivity (e.g., human, mouse, rat) and epitope specificity when selecting antibodies. Validation via knockout cell lines or siRNA knockdown ensures target specificity. Cross-reactivity with other Rel family members (e.g., RelB, c-Rel) should be ruled out. Proper controls, such as nuclear/cytoplasmic fractionation, are critical for interpreting subcellular localization studies. Overall, NF-κB p65 antibodies are indispensable for dissecting the molecular mechanisms of NF-κB-mediated cellular responses.