Phospho-BRAF (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72]
cat.: ET1701-20
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IP, IHC-P
Clonality: Monoclonal
Clone number: JJ08-72
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 84 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Thr401 of Human BRAF aa 371-420 / 766.
Positive control: Mouse testis tissue lysates, PC-12 cell lysates, PC-3M, MCF-7, PC-12, human testis tissue, mouse testis tissue, rat testis tissue, mouse brain tissue, rat brain tissue.
Subcellular location: Nucleus, Cytoplasm, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IP
  IHC-P
1:2,000
1:100
Use at an assay dependent concentration.
1:200
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
ET1701-20_1.jpg Fig1: Western blot analysis of Phospho-BRAF (T401) on different lysates with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/2,000 dilution.

Lane 1: C6 cell lysate
Lane 2: C6 starved overnight then treated with 200nM TPA for 4 hours cell lysate
Lane 3: C6 starved overnight then treated with 200nM TPA for 4 hours cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 3 minutes; ECL: K1801;

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 (ET1701-20) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-20_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1701-20) at 1/200 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.
ET1701-20_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1701-20) at 1/200 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.
ET1701-20_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1701-20) at 1/200 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.
ET1701-20_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1701-20) at 1/200 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.
ET1701-20_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1701-20) at 1/200 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.
ET1701-20_7.jpg Fig7: ICC staining of Phospho-BRAF (T401) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-20_8.jpg Fig8: ICC staining of Phospho-BRAF (T401) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-20_9.jpg Fig9: ICC staining of Phospho-BRAF (T401) in PC-12 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.