Anti-Phospho-NF-kB p65/RelA (Ser468) Rabbit Monoclonal Antibody for WB, IP, ELISA - Q04206

Core Product Specifications and Parameters

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

• Transcription factor p65 (RelA) is the p65 subunit of the NF-kB transcription factor complex, a pleiotropic factor present in almost all cell types, and serves as the endpoint for signal transduction pathways initiated by diverse stimuli related to inflammation, immunity, differentiation, cell growth, tumorigenesis, and apoptosis.
• NF-kB functions as a homo- or heterodimer of Rel domain-containing proteins. The most abundant form is the RELA-NFKB1 (p65-p50) heterodimer. These dimers bind kappa-B DNA sites with distinct affinities, with RELA-NFKB1 and RELA-REL complexes acting as transcriptional activators.
• NF-kB activity is regulated through multiple mechanisms including post-translational modifications, subcellular compartmentalization, and interactions with cofactors or corepressors. Inactive NF-kB is retained in the cytoplasm by I-kB inhibitor proteins; upon IKK-mediated phosphorylation and degradation of I-kB, the NF-kB complex is released and translocates to the nucleus.
• RELA possesses weak intrinsic DNA-binding affinity but contributes to the NF-kB complex's DNA-binding capability. It also modulates chromatin accessibility at target promoters, indirectly influencing gene expression.
• RELA is essential for cytokine gene expression in T-cells, playing a pivotal role in adaptive immune responses. Related references: PMID:15790681
• During SARS-CoV-2 infection, RELA acts as a key transcription factor regulating the interferon response, underscoring its role in antiviral innate immunity. Related references: PMID:33440148
• Subcellularly, RELA shuttles between the cytoplasm and the nucleus. It is subject to extensive post-translational modifications, including acetylation, methylation, phosphorylation (for instance, at Ser468 as targeted by this antibody), S-nitrosylation, and ubiquitin-like conjugation, which modulate its activity, stability, and interactions.

Experimental Guidance and Technical Tips

• For Western blot (WB), prepare lysates using RIPA buffer supplemented with protease and phosphatase inhibitors to preserve the phosphorylation signal at Ser468. Perform SDS-PAGE and transfer, then probe with the antibody at an empirically determined dilution.
• In immunoprecipitation (IP) experiments, confirm that the target protein expression level is sufficient in your cell or tissue lysate. The antibody's rabbit origin and monoclonal nature may facilitate clean capture in species-matching secondary reagents.
• For ELISA-based detection, consider using the antibody as a capture or detection reagent, validating linearity and specificity in your assay system with appropriate phospho-specific controls.
• Since the antibody cross-reacts with human, mouse, and rat, it can be used for comparative studies across these species. However, always verify performance by including positive and negative controls for each species and treatment condition.
• Optimize antibody concentration for each application and sample type, as the recommended dilutions may vary with experimental conditions and detection methods.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your integrated partner for IVD raw materials, providing diagnostic manufacturers and research institutions with a comprehensive portfolio from lead candidate validation antibodies to bulk ancillary reagents. In the field of inflammation and immunology research, we source optimized antibodies, including phospho-specific monoclonal antibodies like this anti-RelA (pS468) product, to accelerate assay development. Our supply chain expertise ensures consistent quality and on-time delivery of critical rare-target reagents to support your clinical and R&D programs.