Phospho-B-Raf-S728 Rabbit pAb for WB - P15056

Core Product Specifications and Parameters

Biological Background: Serine/threonine-protein kinase B-raf Function and Localization

• B-Raf is a serine/threonine-protein kinase that transduces mitogenic signals from the cell membrane to the nucleus, functioning upstream of the MAP kinase cascade by phosphorylating MAP2K1. Related references: PMID:21441910 PMID:29433126
• It also phosphorylates PFKFB2, linking to metabolic regulation. Related references: PMID:36402789
• B-Raf may play a role in the postsynaptic responses of hippocampal neurons. Related references: PMID:1508179
• The protein localizes to the nucleus, cytoplasm, and cell membrane.
• B-Raf is predominantly expressed in the brain and testis.
• It is a proto-oncogene and features multiple regulatory domains, including a zinc-finger and ATP-binding site; it is regulated by phosphorylation, acetylation, and ubiquitination.
• Mutations in BRAF are associated with cardiomyopathies, deafness, ectodermal dysplasia, and intellectual disability.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide around Ser728; the antibody is expected to specifically detect phosphorylated B-Raf at this site.
• For Western blot, consider using phosphatase inhibitors during sample preparation to preserve phosphorylation.
• In ELISA, ensure the coating antigen presents the phosphorylated epitope appropriately.
• Validate antibody specificity using positive controls (e.g., stimulated cell lysates) and negative controls (e.g., phosphatase-treated samples).

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