Anti-Phospho-Raf1 (S259) Rabbit Polyclonal Antibody for WB, IP, ELISA - P04049

Core Product Specifications and Parameters

Biological Background: RAF1 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 signaling cascade, governing cell fate decisions such as proliferation, differentiation, apoptosis, survival, and oncogenic transformation.
• Upon activation, RAF1 initiates a mitogen-activated protein kinase (MAPK) cascade through sequential phosphorylation of dual-specificity MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2).
• RAF1 phosphorylates multiple downstream targets, including BAD (at Ser-75), adenylyl cyclases (ADCY2, ADCY5, ADCY6), PPP1R12A (inhibiting phosphatase activity), and TNNT2 (cardiac muscle troponin T), thereby modulating apoptosis, cAMP signaling, and cardiac function.
• The protein can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation, and angiogenesis (RB1), highlighting its broad regulatory roles.
• RAF1 has been implicated in oncogenic transformation of epithelial cells by upregulating the transcriptional repressor SNAI2/SLUG, which downregulates the tight junction protein occludin (OCLN), and it restricts caspase activation in response to Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
• Subcellularly, RAF1 is found in the cytoplasm, cell membrane, mitochondrion, and nucleus, with translocation to the mitochondria enabling binding to BCL2 and displacement of BAD to protect cells from apoptosis.
• The protein contains a zinc-finger domain, is subject to phosphorylation (including at Ser259, which this antibody recognizes), and plays roles in wound healing, Rho signaling, and migration.
• In skeletal muscle, isoform 1 is more abundant than isoform 2; RAF1 is associated with diseases such as cardiomyopathy and deafness (CMD1NN).

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide spanning Ser259 of human RAF1. This design theoretically targets the phosphorylated epitope; however, users should validate phospho-specificity by comparing signals between phosphatase-treated and untreated lysates in their experimental system.
• Since the antibody is raised in rabbit against a human peptide, cross-reactivity with other species is not guaranteed; validate in the relevant sample system if non-human models are used.
• For Western blot (WB), a band around 73 kDa is expected. Use appropriate blocking and washing conditions to minimize non-specific binding, and include a positive control (e.g., stimulated cell lysates) to confirm phospho-specific detection.
• In immunoprecipitation (IP) and ELISA, the antibody may perform as indicated; consider optimizing antibody concentrations and buffer conditions for your specific assay.
• Avoid repeated freeze-thaw cycles; aliquot and store at -20°C or as recommended.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your one-stop sourcing partner for IVD raw materials, offering validated antibody pairs, optimized monoclonal and polyclonal antibodies, and bulk ancillary reagents to support your RAF1-related diagnostic and research projects. Our comprehensive supply chain also facilitates sourcing of rare-target antibodies and custom raw materials, bridging the gap from concept to clinic.