Anti-Phospho-NF-kB p65/RelA-S536 Rabbit Polyclonal Antibody for WB, IF/ICC, ELISA - Q04206

Core Product Specifications and Parameters

Biological Background: Transcription factor p65 Function and Localization

• NF-kappa-B p65 subunit (RELA) is a central transcription factor of the NF-kappa-B family, encoding a 60 kDa protein (also known as p65, NFKB3, CMCU, AIF3BL3) with critical roles in immune and stress responses. UniProt entry Q04206.
• The NF-kappa-B complex exists as homo- or heterodimers; the RELA-NFKB1 (p50) heterodimer is the most abundant and functions as a transcriptional activator by binding specific κB DNA motifs in gene promoters.
• Activation of NF-kappa-B involves stimulus-induced phosphorylation and degradation of IκB inhibitors, freeing the RELA complex to translocate from the cytoplasm to the nucleus. Phosphorylation of RELA at Ser536, located in the transactivation domain, enhances its transcriptional activity and is a hallmark of canonical NF-κB activation.
• Subcellular localization of RELA is tightly controlled: in resting cells it is predominantly cytoplasmic bound to IκB; upon induction it accumulates in the nucleus to modulate target gene expression. Immunofluorescence can detect nuclear translocation as a readout of pathway activation.
• RELA is heavily modified by phosphorylation, acetylation, methylation, S-nitrosylation, and ubiquitination, which fine-tune its transcriptional output. It is essential for cytokine expression in T-cells (PubMed:15790681) and the type I interferon response during SARS-CoV-2 infection (PubMed:33440148). Related references: PMID:15790681 PMID:33440148
• The RELA gene encodes multiple isoforms, and its activity is modulated through interactions with cofactors and chromatin remodelers, highlighting complex regulatory layers beyond simple nuclear entry.

Experimental Guidance and Technical Tips

• Prepare cell or tissue lysates with protease and phosphatase inhibitor cocktails to preserve phosphorylation at Ser536. For Western blotting, separate proteins by SDS-PAGE and probe with this antibody at a starting dilution verified by the end user; a strong band near 60 kDa is expected.
• For immunofluorescence or immunocytochemistry (IF/ICC), fix cells with 4% paraformaldehyde and permeabilize adequately; use an appropriate blocking buffer to minimize non-specific binding. Investigate nuclear translocation of NF-kB p65 under inflammatory stimuli (e.g., TNF-α treatment) to validate the phospho-specific signal.
• In ELISA applications, consider using a phosphorylated peptide blocking control to confirm phosphorylation-specific detection. Titrate the antibody to determine optimal concentrations in your specific assay context.
• Phosphorylation dynamics can vary across cell types; always include positive and negative controls (e.g., stimulated vs. unstimulated cells) to interpret results accurately.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, laboratories, and research institutes with a one-stop sourcing solution for high-quality IVD raw materials, technical services, and consulting—covering every stage from concept to clinic. This anti-Phospho-NF-kB p65/RelA-S536 rabbit polyclonal antibody exemplifies our commitment to supplying premium immunoassay reagents for the NF-kB signaling pathway, relevant to inflammation, oncology, and immunology research. Beyond single antibodies, we offer validated antibody pairs, optimized recombinant proteins, and ancillary critical raw materials—all designed to streamline your assay development.