Anti-Phospho-NF-kB p65/RelA-T435 Rabbit Polyclonal Antibody for WB and ELISA - Q04206

Core Product Specifications and Parameters

Biological Background: NF-kB p65/RelA Function and Localization

• Encoded by the RELA gene, NF-kB p65 (also known as RelA) is a subunit of the NF-kB transcription factor complex, which plays a central role in regulating immune responses, inflammation, cell proliferation, and apoptosis.
• NF-kB functions as a homo- or heterodimer; the RELA-NFKB1 heterodimer is the most abundant form, and the RELA-RELA homodimer can also activate transcription at specific target gene promoters.
• In unstimulated cells, NF-kB dimers are sequestered in the cytoplasm by inhibitor of kB (IkB) proteins; upon stimulation, IkB is phosphorylated by IKK and degraded, allowing NF-kB to translocate to the nucleus and activate gene expression.
• RELA is essential for cytokine gene expression in T-cells, contributing to adaptive immune responses. Related references: PMID:15790681
• It has been identified as a key transcription factor regulating the interferon response during SARS-CoV-2 infection. Related references: PMID:33440148
• RELA is subject to extensive post-translational modifications, including phosphorylation (e.g., at Thr435), acetylation, methylation, and others, which modulate its transcriptional activity and interactions.
• The protein localizes primarily to the cytoplasm in its inactive state and to the nucleus upon activation, where it binds DNA and recruits coactivators.
• Alternative splicing and chromosomal rearrangements involving RELA have been associated with certain diseases, highlighting its importance in both normal physiology and pathology.

Experimental Guidance and Technical Tips

• The immunogen used to generate this antibody is a synthetic phosphorylated peptide around Thr435, so it specifically recognizes the phosphorylated form of p65. For WB, ensure that cell lysates are prepared with phosphatase inhibitors to preserve the phosphorylation state.
• When performing ELISA, consider using both phosphorylated and non-phosphorylated peptides to verify the antibody's phospho-specificity.
• For western blotting, a band around 60 kDa is expected; validate using positive controls known to have activated NF-kB (e.g., TNF-alpha-treated cells) and negative controls (e.g., untreated or phosphatase-treated lysates).
• The antibody is reported to react with human and mouse samples; cross-reactivity with other species should be evaluated experimentally if needed.
• Optimize primary antibody dilutions based on your sample system; typical starting points are 1:500-1:2000 for WB, but exact conditions may vary.

CamelBio: Your One-Stop Sourcing Bridge

• CamelBio serves as a comprehensive IVD raw material partner, supplying high-quality antibodies, antigens, and ancillary reagents for diagnostic assay development. For NF-kB signaling research, we offer this phospho-specific RELA antibody alongside validated antibody pairs and bulk intermediates to streamline your workflow. With one-stop access from concept to clinic, CamelBio supports your project with technical consulting and reliable sourcing of rare and standard targets alike.