Src Recombinant Rabbit Monoclonal Antibody [JF0947]
cat.: ET1702-03
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JF0947
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: 60 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Src aa 20-60.
Positive control: HUVEC cell lysate, A549 cell lysate, PC-3M cell lysate, SH-SY5Y cell lysate, Wild-type HCT116 whole cell lysates, SK-OV-3, Hela, MCF-7, A431, human tonsil tissue, human breast carcinoma tissue, human kidney tissue, rat brain tissue, mouse kidney tissue, NIH/3T3, NIH/3T3 cell lysate.
Subcellular location: Cell membrane, Mitochondrion inner membrane, Nucleus, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:1,000-1:5,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P12931 Human | P05480 Mouse | Q9WUD9 Rat
Alternative names: ASV Avian sarcoma virus c SRC CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA cSrc EC 2.7.10.2 Neuronal CSRC tyrosine specific protein kinase Neuronal SRC Oncogene SRC OTTHUMP00000174476 OTTHUMP00000174477 p60 Src p60-Src p60Src pp60c src pp60c-src pp60csrc Proto oncogene tyrosine protein kinase Src Proto-oncogene c-Src Proto-oncogene tyrosine-protein kinase Src Protooncogene SRC Protooncogene SRC Rous sarcoma Src SRC Oncogene SRC proto oncogene non receptor tyrosine kinase SRC_HUMAN SRC1 Tyrosine kinase pp60c src Tyrosine protein kinase SRC 1 Tyrosine protein kinase SRC1 v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian
Images
ET1702-03_1.jpg Fig1: All lanes: Western blot analysis of Src with anti-Src antibody [JF0947] (ET1702-03) at 1:1,000 dilution.
Lane 1: Wild-type HCT116 whole cell lysate (20 µg).
Lane 2: Src knockout HCT116 whole cell lysate (20 µg).

ET1702-03 was shown to specifically react with Src in wild-type HCT116 cells. No band was observed when Src knockout sample was tested. Wild-type and Src knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1702-03, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) 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.
ET1702-03_2.jpg Fig2: Western blot analysis of Src 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 (ET1702-03, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: HUVEC cell lysate
Lane 2: A549 cell lysate
Lane 3: PC-3M cell lysate
Lane 4: SH-SY5Y cell lysate
ET1702-03_3.jpg Fig3: Immunocytochemistry analysis of SK-OV-3 cells labeling Src with Rabbit anti-Src antibody (ET1702-03) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Rabbit anti-Src antibody (ET1702-03) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1702-03_4.jpg Fig4: ICC staining of Src 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 (ET1702-03, 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).
ET1702-03_5.jpg Fig5: ICC staining of Src 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 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-03, 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).
ET1702-03_6.jpg Fig6: ICC staining of Src 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 (ET1702-03, 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).
ET1702-03_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Src 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 (ET1702-03, 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.
ET1702-03_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Src 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 (ET1702-03, 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.
ET1702-03_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Src 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 (ET1702-03, 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.
ET1702-03_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Src 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 (ET1702-03, 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.
ET1702-03_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-Src 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 (ET1702-03, 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.
ET1702-03_12.jpg Fig12: Flow cytometric analysis of Src was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-03, 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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1702-03_13.jpg Fig13: Western blot analysis of Src on NIH/3T3 cell lysates with Rabbit anti-Src antibody (ET1702-03) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Exposure time: 2 minutes;

8% 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 (ET1702-03) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.