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Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Cytokeratin 7 antibody (R1309-4) at 1/5,000 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 (R1309-4) at 1/5,000 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.
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Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Cytokeratin 7 antibody (R1309-4) at 1/5,000 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 (R1309-4) at 1/5,000 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.
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☑ Knockdown (KD)
All lanes: Western blot analysis of Cytokeratin 7 with anti-Cytokeratin 7 antibody(R1309-4) at 1:1,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (20 µg).
Lane 3/4: Cytokeratin 7 fragment 1 knockdown Hela whole cell lysate (20 µg).
Lane 5/6: Cytokeratin 7 fragment 2 knockdown Hela whole cell lysate (20 µg).
R1309-4 was shown to specifically react with Cytokeratin 7 in wild-type Hela cells. Weakened bands were observed when Cytokeratin 7 knockdown samples were tested. Wild-type and Cytokeratin 7 knockdown 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 (R1309-4, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were 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.
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Western blot analysis of Cytokeratin 7 on A549 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1/500 dilution 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.
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ICC staining Cytokeratin 7 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 Cytokeratin 7 polyclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining Cytokeratin 7 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 Cytokeratin 7 polyclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining Cytokeratin 7 in SW480 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 Cytokeratin 7 polyclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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Flow cytometric analysis of Cytokeratin 7 was done on Hela cells. The cells were fixed, permeabilized and stained with Cytokeratin 7 antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.
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