Phospho-NF-kB p65/RelA-T505 Rabbit pAb - Q04206

Core Product Specifications and Parameters

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

• NF-kB p65/RelA is a subunit of the NF-kB transcription factor complex, which is a pleiotropic regulator activated by diverse stimuli to control genes involved in inflammation, immunity, cell growth, differentiation, tumorigenesis, and apoptosis.
• The active NF-kB complex can be a homo- or heterodimer of Rel-family proteins; the RELA-p50 heterodimer is the most abundant form, binding to k-B sites in target DNA and acting as a transcriptional activator.
• Other Rel-family members include RELB, NFKB1 (p105/p50), NFKB2 (p52), and c-REL, with distinct dimer combinations providing specificity and functional diversity.
• In resting cells, NF-kB is retained in the cytoplasm through interaction with inhibitor I-kB proteins; upon stimulation, I-kB phosphorylation and degradation releases NF-kB for nuclear translocation.
• RELA is essential for cytokine gene expression in T-cells, a key function in adaptive immunity. Related references: PMID:15790681
• During SARS-CoV-2 infection, RELA serves as a critical transcription factor regulating the interferon response. Related references: PMID:33440148
• The protein undergoes extensive post-translational modifications (e.g., phosphorylation, acetylation, methylation, ubiquitination) that modulate its activity, stability, and interactions.
• Subcellular localization cycles between nucleus and cytoplasm, with phosphorylation at various sites (including Thr505) influencing transcriptional activity or complex dynamics.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide covering Thr505, so antibody reactivity is phosphorylation-dependent. Ensure samples are prepared with phosphatase inhibitors to preserve the phosphorylated epitope.
• For western blot, use standard SDS-PAGE and transfer protocols; the expected band size for phosphorylated RELA is ~60 kDa. Optimize blocking and antibody dilution conditions empirically.
• In ELISA, plate coating with phosphorylated peptide or protein lysate may be used; cross-reactivity with non-phosphorylated forms should be assessed in the experimental system.
• This antibody is reactive with human samples; for other species, sequence homology around Thr505 should be checked before use.

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