Anti-Phospho-Raf1 (Ser621) Rabbit Polyclonal Antibody - 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 encoded by the RAF1 gene. It functions as a critical regulatory link between membrane-associated Ras GTPases and the MAPK/ERK signaling cascade, controlling key cellular processes such as proliferation, differentiation, apoptosis, and oncogenic transformation.
• Activated RAF1 phosphorylates and activates the dual-specificity kinases MAP2K1/MEK1 and MAP2K2/MEK2, which in turn phosphorylate the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, propagating downstream signals.
• Beyond the canonical MAPK pathway, RAF1 phosphorylates multiple other substrates, including BAD (at Ser-75), adenylyl cyclases (ADCY2, ADCY5, ADCY6), PPP1R12A, and TNNT2, linking it to diverse cellular functions such as apoptosis regulation and cAMP signaling.
• RAF1 localizes to the cytoplasm, cell membrane, mitochondrion, and nucleus. Its translocation to mitochondria enables binding to BCL2 and displacement of BAD, providing an additional anti-apoptotic mechanism.
• In skeletal muscle, isoform 1 of RAF1 is more abundant than isoform 2, suggesting tissue-specific regulatory roles.
• RAF1 is a proto-oncogene implicated in various cancers; its dysregulated activity promotes oncogenic transformation, in part by repressing tight junction proteins like occludin through upregulation of SNAI2/SLUG.
• The protein contains a zinc-finger domain and is subject to multiple post-translational modifications, including phosphorylation, which tightly regulate its kinase activity and subcellular distribution.

Experimental Guidance and Technical Tips

• This antibody was raised against a synthetic phosphorylated peptide corresponding to Ser621 of human RAF1. It is expected to specifically recognize phosphorylated RAF1 at this residue. For total RAF1 detection, a pan-antibody may be required as a control.
• For Western blot, use standard protocols but ensure phosphatase inhibitors are included in lysis and incubation buffers to preserve phosphorylation. Detection of a band near 73 kDa is anticipated in appropriate human samples.
• For immunoprecipitation, optimize antibody concentration and incubation time for your experimental system. Pre-clearing and appropriate isotype controls are recommended.
• For ELISA, this antibody can be used for capture or detection; validate binding affinity and specificity with phosphorylated and non-phosphorylated peptide controls.
• Always validate the antibody in your specific sample matrix and experimental conditions; performance can vary with cell type and stimulation status.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, laboratories, and research institutes with comprehensive access to IVD raw materials, including validated antibody pairs, optimized polyclonal and monoclonal antibodies, and ancillary reagents for assay development. For researchers investigating the MAPK pathway or oncogenic signaling, we offer reliable antibodies like this phospho-specific RAF1 (Ser621) polyclonal antibody, along with bulk supply and tailored support to streamline your project from concept to clinical application. Let CamelBio be your sourcing bridge for all your IVD raw material needs.