NCK1 Recombinant Rabbit Monoclonal Antibody [JU41-80]
cat.: ET7106-57
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JU41-80
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: 43 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human NCK1 aa 204-296 / 377.
Positive control: Hela cell lysate, NIH/3T3 cell lysate, rat kidney tissue lysate, mouse testis tissue lysate, LOVO, A431, HUVEC, mouse testis tissue, rat testis tissue, human tonsil tissue, human kidney tissue, Jurkat.
Subcellular location: Cytoplasm. Endoplasmic reticulum. Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:100
1:10-1:50
Uniprot #: SwissProt: P16333 Human | Q99M51 Mouse | B2RZ33 Rat
Alternative names: Cytoplasmic protein NCK1 Melanoma Nck protein MGC12668 NCK 1 NCK adaptor protein 1 NCK alpha NCK NCK tyrosine kinase Nck-1 NCK1 NCK1_HUMAN NCKalpha Non catalytic region of tyrosine kinase SH2/SH3 adaptor protein NCK alpha SH2/SH3 adaptor protein NCK-alpha
Images
ET7106-57_1.jpg Fig1: Western blot analysis of NCK1 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 (ET7106-57, 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: NIH/3T3 cell lysate
Lane 3: Rat kidney tissue lysate
Lane 4: Mouse testis tissue lysate

Predicted band size: 43 kDa
Observed band size: 43/36 kDa
ET7106-57_2.jpg Fig2: ICC staining of NCK1 in LOVO 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 (ET7106-57, 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).
ET7106-57_3.jpg Fig3: ICC staining of NCK1 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 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-57, 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).
ET7106-57_4.jpg Fig4: ICC staining of NCK1 in HUVEC 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 (ET7106-57, 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).
ET7106-57_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 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.
ET7106-57_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 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.
ET7106-57_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 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.
ET7106-57_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 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.
ET7106-57_9.jpg Fig9: Flow cytometric analysis of NCK1 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7106-57, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-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).
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