Anti-B-Raf (V600E) Mouse Monoclonal Antibody, 100 µl (26039)

Background B-Raf is a member of the Raf family of Ser/Thr protein kinases. It functions downstream of Ras to regulate the MAP kinase signaling pathway. Mutations in the BRAF gene cause diseases. Inherited mutations in BRAF cause cardiofaciocutaneous syndrome. Acquired mutations in BRAF have been found in cancers. Tested Applications ELISA, WB, IHC, IP Source Species Mouse Species Reactivity B-Raf(V600E) antibody recognizes mutated, but not wild-type, B-Raf(V600E) proteins. Datasheet Validation Data: Western Blot Western blot analysis of recombinant B-raf proteins. Purified His-tagged B-Raf(V600E) (amino acids 513-693, lane 2) and corresponding wild type protein (lane 1) were blotted with anti-B-Raf(V600E) monoclonal antibody (Cat. #NEBA10126). Immunohistochemistry Immunohistochemical analysis of paraffin-embedded human melanoma tissue with anti-B-Raf (V600E) monoclonal antibody (Cat. #NEBA10126). Tissue samples were fixed with formaldehyde and blocked with 1% serum for 15 min at 37 C. Antigen retrieval was by heat mediation in citrate buffer (pH6). Samples were then incubated with primary antibody (1:100) overnight at 4C. A HRP-conjugated Goat anti-mouse IgG (dilution 1:50) was used as secondary antibody. Immunofluorescence Immunofluorescence of cells expressing B-Raf (V600E) protein with anti-B-Raf (V600E) antibody. HEK293T cells were transfected with pCDNA3-GFP-B-Raf WT plasmid (left column) or pCDNA3-GFP-B-Raf V600E plasmid (right column), then fixed and stained with anti-B-Raf (V600E) monoclonal antibody (Cat. #NEBA10126). Application data from publications: Positive mutant specific stain on primary (b) and metastasized (d) melanoma. a, b are H&E stain of primary and metastasized melanoma (Ref 1) BRAF mutant (V600E) specific IHC stain on confrimed mutant containing primary melanoma samples (Ref 2) Recent publications using Anti-B-Raf (V600E): Immunohistochemistry with a mutation-specific monoclonal antibody as a screening toll for BRAFV600E mutational status in primary cutaneous malignant melanoma. JK Feller, S Yang and M Mahalingam. Modern Pathology (2013) 26, 414-420 Immunohistochemistry as a quick screening method for clinical detection of BRAF (V600E) mutation in melanoma patients. Q Chen, X Xia et al. Tumor Biol. (2013) DOI 10.1007/s13277-014-1759-6 DNA vaccine elicits an efficient antitumor response by targeting the mutant Kras in a transgenic mouse lung cancer model. TY Weng, MC Yen, CT Huang, JJ Hung, YL Chen. Gene Ther. 2014 Jul 31. doi: 10.1038/gt.2014.67