A-RAF Recombinant Rabbit Monoclonal Antibody [JJ08-68]
cat.: ET1701-18
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JJ08-68
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: 68 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human A-RAF aa 146-310 / 606.
Positive control: Hela cell lysate, A431 cell lysate, Hela, A431, NIH/3T3, human kidney tissue, human breast carcinoma tissue.
Subcellular location: Cytosol.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P10398 Human | P04627 Mouse
Alternative names: A raf 1 A Raf proto oncogene serine/threonine protein kinase ARAF 1 Araf ARaf proto oncogene serine/threonine protein kinase ARAF_HUMAN ARAF1 Oncogene Araf1 Oncogene PKS2 PKS 2 PKS PKS2 Proto oncogene Pks Proto-oncogene A-Raf Proto-oncogene A-Raf-1 Proto-oncogene Pks RAFA 1 RAFA1 Ras binding protein DA Raf Serine/threonine-protein kinase A-Raf v raf murine sarcoma 3611 viral oncogene homolog v raf murine sarcoma 3611 viral oncogene homolog 1 v raf oncogene homolog 1 (murine sarcoma 3611 virus)
Images
ET1701-18_1.jpg Fig1: Western blot analysis of A-RAF on different lysates with Rabbit anti-A-RAF antibody (ET1701-18) at 1/1,000 dilution.

Lane 1: A549-WT cell lysate
Lane 2: A549-KD A-RAF cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 68 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 (ET1701-18) at 1/1,000 dilution was used in 2% 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.
ET1701-18_2.jpg Fig2: Western blot analysis of A-RAF on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-18, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: A431 cell lysate
ET1701-18_3.jpg Fig3: ICC staining of A-RAF in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-18_4.jpg Fig4: ICC staining of A-RAF in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-18_5.jpg Fig5: ICC staining of A-RAF in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-18, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-18_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-A-RAF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-18, 1/50) for 30 minutes 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.
ET1701-18_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-A-RAF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-18, 1/50) for 30 minutes 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.
ET1701-18_8.jpg Fig8: Flow cytometric analysis of A-RAF was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-18, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.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”.