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B-Raf (V600E)

Catalog Number:CR-M 26039

Gene Symbol: BRAF, BRAF1, RAFB1

Description: Anti-B-Raf (V600E) Mouse Monoclonal Antibody

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

Immunogen: A synthetic peptide from the internal region of Braf (V600E), human origin.

Tested applications: ELISA, WB, IHC, IP

Recommended dilutions:

ELISA 1:1000-1:5000

WB 1:500-1:2000

IHC 1:100-1:200

Concentration: 1 mg/ml

Host: Mouse

Clonality: Monoclonal

Isotype: IgG

Purity: Purified from ascites

Format: Liquid Storage buffer:

Preservative: no

Constituents: PBS (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 50% glycerol

Species Reactivity: Recognizes V600E mutated, but not wild-type B-Raf proteins from vertebrates.

Storage Conditions: Store at -20°C. Avoid freeze / thaw cycles.

Western blot:

Western blot analysis of recombinant B-raf proteins. Purified His-tagged B-Raf(V600E) protein (amino acids 513-693, lane 2) and corresponding wild type protein (lane 1) were blotted with anti-B-Raf(V600E) monoclonal antibody (Cat. # 26039).

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. # 26039).

Immunohistochemistry:

Immunohistochemical analysis ofparaffin-embedded human melanoma tissuewith anti-B-Raf (V600E) monoclonal antibody (Cat. # 26039). Tissue samples were fixed with formaldehyde and blocked with 1% serum for 15 min at 37 °C. Antigen retrieval was by heatmediation in citrate buffer (pH6). Samples were then incubated with primary antibody (1:100)overnight at 4°C. A HRP-conjugated Goatanti-mouse IgG (dilution 1:50) was used as secondary antibody.

Immunohistochemical analysis of paraffinembedded Thyroid Carcinoma tissue -withantiBRaf(V600E) monoclonal antibody (Cat. # 26039 ). Tissue samples were fixed withparaffin.

Samples were then incubated with primaryantibody (1:100) overnight at 4°C. A HRP-conjugated Goat anti-mouse IgG (dilution 1:50) was used as secondary antibody.Allele specific PCR validated to be Negative for BRAF V600E.

For research use only. Not for diagnostic or therapeutic applications.

Immunohistochemical analysis of paraffin embedded Thyroid Carcinoma tissue -with anti BRaf(V600E) monoclonal antibody

(Cat. # 26039). Tissue samples were fixed withparaffin. Samples were then incubated with primary antibody (1:100) overnight at 4°C. A HRP-conjugated Goat anti-mouse IgG (dilution 1:50) was used as secondary antibody. Allele specific PCR validated to be Negative for BRAF V600E.

Publications:

1. John K Feller, Shi Yang and Meera Mahalingam. Immunohistochemistry with a mutation-specific

monoclonal antibody as a screening tool for the

BRAFV600E mutational status in primary cutaneous malignant melanoma. Modern Pathology. 2012.doi:10.1038/modpathol.2012.168