Anti-Phospho-Raf1-S621 Rabbit Monoclonal Antibody for WB, ELISA - P04049

Core Product Specifications and Parameters

Biological Background: RAF1 (c-Raf) Function and Localization

• RAF1 (also known as c-Raf or Raf-1) is a serine/threonine-protein kinase that acts as a critical regulatory link between membrane-associated Ras GTPases and the MAPK/ERK cascade, functioning as a switch controlling cell fate decisions including proliferation, differentiation, apoptosis, and survival.
• The kinase initiates a mitogen-activated protein kinase (MAPK) cascade by sequentially phosphorylating the dual-specificity MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2).
• RAF1 phosphorylates diverse substrates: BAD (at Ser-75, via PAK1), adenylyl cyclases (ADCY2, ADCY5, ADCY6), PPP1R12A (inhibiting phosphatase activity), and TNNT2; it also promotes NF-κB activation and inhibits ROCK2, ASK1, and MST2 kinases.
• The protein can protect cells from apoptosis by translocating to mitochondria, where it binds BCL2 and displaces BAD; it also restricts caspase activation in response to Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
• RAF1 contributes to oncogenic transformation through repression of the tight junction protein occludin (OCLN) by inducing up-regulation of the transcriptional repressor SNAI2/SLUG.
• Subcellular localization includes cytoplasm, cell membrane, mitochondrion, and nucleus, reflecting its multifaceted regulatory roles.
• The protein features a zinc-finger domain, binds ATP, and is subject to phosphorylation (e.g., S621), methylation, and alternative splicing (isoform 1 is more abundant in skeletal muscle).
• Keywords associated with RAF1 include cardiomyopathy, deafness, disease variant, proto-oncogene, and serine/threonine-protein kinase, indicating its involvement in both normal physiology and disease.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide; ensure samples are processed with phosphatase inhibitors to preserve phosphorylation status during lysis and preparation.
• For Western blotting, expect a band at approximately 73 kDa; include appropriate controls such as phosphatase-treated lysates or stimulated versus unstimulated cells to confirm phospho-specificity.
• Since RAF1 phosphorylation is dynamic, consider using a total RAF1 antibody in parallel to normalize signals and accurately interpret activation changes.
• In ELISA, optimize the coating antigen concentration and antibody dilution; a standard curve with a known phospho-peptide may aid quantification.
• Cross-reactivity with rat tissue is reported; validate the antibody’s performance in your specific rat model system, as signal intensity may vary across samples.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, labs, and research institutes with one-stop access to IVD raw materials, technical services, and consulting—covering every stage from concept to clinic. For studies focused on the RAS-RAF-MEK-ERK signaling axis, we offer a comprehensive portfolio including validated antibody pairs, optimized monoclonal and polyclonal antibodies, bulk ancillary reagents, and rare target raw-material sourcing to accelerate your assay development.