B Raf (BRAF) Mouse Monoclonal Antibody [Clone ID: OTI5A9]

CAT#: CF500431

Carrier-free (BSA/glycerol-free) BRAF mouse monoclonal antibody, clone OTI5A9 (formerly 5A9)

Product Images

HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY BRAF (RC211013, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-BRAF. Positive lysates LY401382 (100ug) and LC401382 (20ug) can be purchased separately from OriGene.

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  Western blot analysis of extracts (35ug) from 9 different cell lines by using anti-BRAF monoclonal antibody.

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  Western blot analysis of extracts (35ug) from 4 tissue lysates by using anti-BRAF monoclonal antibody (1:500).

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  Equivalent amounts of cell lysates (10 ug per lane) of wild-type HeLa cells (WT, Cat# LC810HELA) and BRAF-Knockout HeLa cells (KO, Cat# LC835315) were separated by SDS-PAGE and immunoblotted with anti-BRAF monoclonal antibody TA500431 (1:500). Then the blotted membrane was stripped and reprobed with anti-PCNA antibody as a loading control.

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  Immunohistochemical staining of paraffin-embedded Adenocarcinoma of Human colon tissue using anti-BRAF mouse monoclonal antibody. (TA500431)

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  Immunohistochemical staining of paraffin-embedded Carcinoma of Human thyroid tissue using anti-BRAF mouse monoclonal antibody. (TA500431)

• Immunohistochemical staining of paraffin-embedded Adenocarcinoma of Human endometrium tissue using anti-BRAF mouse monoclonal antibody. (TA500431)

• Immunohistochemical staining of paraffin-embedded Human prostate tissue within the normal limits using anti-BRAF mouse monoclonal antibody. (TA500431)

• Immunohistochemical staining of paraffin-embedded Carcinoma of Human prostate tissue using anti-BRAF mouse monoclonal antibody. (TA500431)

• Immunohistochemical staining of paraffin-embedded Human bladder tissue within the normal limits using anti-BRAF mouse monoclonal antibody. (TA500431)

• Immunohistochemical staining of paraffin-embedded Human lymphoma tissue using anti-BRAF mouse monoclonal antibody. (TA500431)

• Anti-BRAF mouse monoclonal antibody (TA500431) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY BRAF (RC211013).

• Immunoprecipitation (IP) of BRAF by using TrueMab monoclonal anti-BRAF antibodies (Negative control: IP without adding anti-BRAF antibody.). For each experiment, 500ul of DDK tagged BRAF overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of anti-BRAF antibody and 20ul (0.1mg) of goat anti-mouse conjugated magnetic beads were mixed and incubated overnight. After extensive wash to remove any non-specific binding, the immuno-precipitated products were analyzed with rabbit anti-DDK polyclonal antibody.

• HEK293T cells transfected with either RC211013 overexpress plasmid (Red) or empty vector control plasmid (Blue) were immunostained by anti-BRAF antibody (TA500431), and then analyzed by flow cytometry.

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Specifications

Product Data

• Clone Name: OTI5A9
• Applications: FC, IF, IHC, IP, WB
• Recommended Dilution: WB 1:1000~2000, IF 1:100, FLOW 1:100
• Reactivities: Human, Monkey, Mouse, Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Full length human recombinant protein of human BRAF (NP_004324) produced in HEK293T cell.
• Formulation: Lyophilized powder (original buffer 1X PBS, pH 7.3, 8% trehalose)
• Reconstitution Method: For reconstitution, we recommend adding 100uL distilled water to a final antibody concentration of about 1 mg/mL. To use this carrier-free antibody for conjugation experiment, we strongly recommend performing another round of desalting process. (OriGene recommends Zeba Spin Desalting Columns, 7KMWCO from Thermo Scientific)
• Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G)
• Conjugation: Unconjugated
• Storage: Store at -20°C as received.
• Stability: Stable for 12 months from date of receipt.
• Predicted Protein Size: 84.4 kDa
• Gene Name: Homo sapiens B-Raf proto-oncogene, serine/threonine kinase (BRAF), transcript variant 1, mRNA.
• Database Link:
  NP_004324
  Entrez Gene 109880 MouseEntrez Gene 114486 RatEntrez Gene 693554 MonkeyEntrez Gene 673 Human
  UniProt ID P15056
• Background: 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.
• Synonyms: B-raf; B-RAF1; BRAF1; NS7; RAFB1

Reference Data

• Protein Families: Druggable Genome, Protein Kinase
• Protein Pathways: Acute myeloid leukemia, Bladder cancer, Chemokine signaling pathway, Chronic myeloid leukemia, Colorectal cancer, Endometrial cancer, ErbB signaling pathway, Focal adhesion, Glioma, Insulin signaling pathway, Long-term depression, Long-term potentiation, MAPK signaling pathway, Melanoma, mTOR signaling pathway, Natural killer cell mediated cytotoxicity, Neurotrophin signaling pathway, Non-small cell lung cancer, Pancreatic cancer, Pathways in cancer, Progesterone-mediated oocyte maturation, Prostate cancer, Regulation of actin cytoskeleton, Renal cell carcinoma, Thyroid cancer, Vascular smooth muscle contraction