BRAF Mouse Monoclonal Antibody [F1-F4-E10]
cat.: EM1701-32
Product Type: Mouse monoclonal IgG2b, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: F1-F4-E10
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 84 kDa
Isotype: IgG2b
Immunogen: Recombinant protein within Human BRAF aa 49-239 / 766.
Positive control: HeLa cell lysate, K-562 cell lysate, HT-29 cell lysate, SK-MEL-28 cell lysate, HEK-293 cell lysate, SiHa cell lysate, COS-1 cell lysate, Vero cell lysate, PC-12 cell lysate, HAP1 cell lysate, rat testis tissue, human testis tissue, human colon cancer tissue, K-562.
Subcellular location: Nucleus, Cytoplasm, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:100
1:500-1:1,000
1:1,000
Uniprot #: SwissProt: P15056 Human | P28028 Mouse
Entrez Gene: 114486 Rat
Alternative names: FLJ95109 94 kDa B raf protein B raf 1 B Raf proto oncogene serine threonine protein kinase B Raf proto oncogene, serine/threonine kinase B RAF1 B-Raf proto-oncogene serine/threonine-protein kinase (p94) BRAF 1 BRAF BRAF_HUMAN BRAF1 cRmil MGC126806 MGC138284 Murine sarcoma viral (v-raf) oncogene homolog B1 Murine sarcoma viral v raf oncogene homolog B1 NS7 Oncogen BRAF oncogene BRAF1 p94 Proto-oncogene B-Raf Proto-oncogene c-Rmil RAFB 1 RAFB1 RMIL Serine/threonine-protein kinase B-raf v raf murine sarcoma viral oncogene homolog B v raf murine sarcoma viral oncogene homolog B1 v-Raf murine sarcoma viral oncogene homolog B1
Images
EM1701-32_1.jpg Fig1: Western blot analysis of BRAF on different lysates with Mouse anti-BRAF antibody (EM1701-32) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: K-562 cell lysate (20 µg/Lane)
Lane 3: HT-29 cell lysate (20 µg/Lane)
Lane 4: SK-MEL-28 cell lysate (20 µg/Lane)
Lane 5: HEK-293 cell lysate (20 µg/Lane)
Lane 6: SiHa cell lysate (20 µg/Lane)
Lane 7: COS-1 cell lysate (20 µg/Lane)
Lane 8: Vero cell lysate (20 µg/Lane)
Lane 9: PC-12 cell lysate (20 µg/Lane)

Predicted band size: 84 kDa
Observed band size: 84 kDa

Exposure time: 3 minutes;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-32) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1701-32_2.jpg Fig2: Western blot analysis of BRAF on different lysates with Mouse anti-BRAF antibody (EM1701-32) at 1/5,000 dilution.

Lane 1: HAP1-parental cell lysate (10 µg/Lane)
Lane 2: HAP1-BRAF KD cell lysate (10 µg/Lane)

Predicted band size: 84 kDa
Observed band size: 84 kDa

Exposure time: 30 seconds; ECL: K1802;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-32) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1701-32_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-BRAF antibody (EM1701-32) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1701-32) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM1701-32_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-BRAF antibody (EM1701-32) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1701-32) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM1701-32_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-BRAF antibody (EM1701-32) at 1/500 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1701-32) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM1701-32_6.jpg Fig6: Immunocytochemistry analysis of K-562 cells labeling BRAF with Mouse anti-BRAF antibody (EM1701-32) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-BRAF antibody (EM1701-32) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
EM1701-32_7.jpg Fig7: Flow cytometric analysis of K-562 cells labeling BRAF.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-32, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.