BRAF (phospho S446) polyclonal antibody
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic phosphopeptide of BRAF.
Immunogen
Synthetic phosphopeptide (conjugated with KLH) corresponding to residues surrounding S446 of human BRAF. This sequence is conserved in B-Rafs from many species, and has high homology to C-Raf (Ser-338) and A-Raf (Ser-299).
Host
Rabbit
Theoretical MW (kDa)
95
Reactivity
Human, Mouse, Rat
Specificity
This antibody was cross-adsorbed to phospho-B-Raf (Ser-579) peptide before affinity purification using phospho-B-Raf (Ser-446) peptide. The purified antibody detects a strong band at 95 kDa* and a weaker band at 105 kDa in western blots of human Jurkat or A431 cells treated with Calyculin A. The antibody also detects other phosphorylated proteins at 150 and 200 kDa in calyculin A treated cells.
*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.Form
Liquid
Purification
Affinity purification
Recommend Usage
ELISA (1:2000)
Western Blot (1:1000) (Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1 hour at room temperature.)
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS (50% glycerol, 1 mg/mL BSA, 0.05% sodium azide).
Storage Instruction
Store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Western blot of human Jurkat cells treated with calyculin A (100 nM) for 30 min. The blots were untreated (lanes 1 & 3) or treated (lanes 2 & 4) with lambda phosphatase and probed with anti-B-Raf (N-terminus) (lanes 1 & 2) or anti-B-Raf (Ser-446) (lanes 3 & 4).Enzyme-linked Immunoabsorbent Assay
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Gene Info — BRAF
Entrez GeneID
673Protein Accession#
P15056Gene Name
BRAF
Gene Alias
B-RAF1, BRAF1, FLJ95109, MGC126806, MGC138284, RAFB1
Gene Description
v-raf murine sarcoma viral oncogene homolog B1
Gene Ontology
HyperlinkGene Summary
This gene encodes a protein belonging to the raf/mil family of serine/threonine protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion. Mutations in this gene are associated with cardiofaciocutaneous syndrome, a disease characterized by heart defects, mental retardation and a distinctive facial appearance. Mutations in this gene have also been associated with various cancers, including non-Hodgkin lymphoma, colorectal cancer, malignant melanoma, thyroid carcinoma, non-small cell lung carcinoma, and adenocarcinoma of lung. A pseudogene, which is located on chromosome X, has been identified for this gene. [provided by RefSeq
Other Designations
94 kDa B-raf protein|B-Raf proto-oncogene serine/threonine-protein kinase (p94)|Murine sarcoma viral (v-raf) oncogene homolog B1
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Interactome
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Pathway
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Disease
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Publication Reference
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Rheb inhibits C-raf activity and B-raf/C-raf heterodimerization.
Karbowniczek M, Robertson GP, Henske EP.
The Journal of Biological Chemistry 2006 Sep; 281(35):25447.
Application:WB-Tr, Human, HEK 293 cells.
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Functional analysis of the regulatory requirements of B-Raf and the B-Raf(V600E) oncoprotein.
Brummer T, Martin P, Herzog S, Misawa Y, Daly RJ, Reth M.
Oncogene 2006 Oct; 25(47):6262.
Application:WB-Tr, Human, HEK 293, MCF10A cells.
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BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis.
Wilhelm SM, Carter C, Tang L, Wilkie D, McNabola A, Rong H, Chen C, Zhang X, Vincent P, McHugh M, Cao Y, Shujath J, Gawlak S, Eveleigh D, Rowley B, Liu L, Adnane L, Lynch M, Auclair D, Taylor I, Gedrich R, Voznesensky A, Riedl B, Post LE, Bollag G, Trail PA.
Cancer Research 2004 Oct; 64(19):7099.
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Rheb inhibits C-raf activity and B-raf/C-raf heterodimerization.
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