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Western blot analysis of Glutathione Peroxidase 1 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 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.
Positive control:
Lane 1: Mouse liver tissue lysate, untreated
Lane 2: Human liver tissue lysate, untreated
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ICC staining Glutathione Peroxidase 1 in 293T 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 antibody (R1401-12) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution.
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Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Glutathione Peroxidase 1 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 antibody (R1401-12) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-Glutathione Peroxidase 1 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 antibody (R1401-12) at 1/50 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Glutathione Peroxidase 1 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 antibody (R1401-12) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Glutathione Peroxidase 1 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 antibody (R1401-12) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Glutathione Peroxidase 1 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 antibody (R1401-12) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of Glutathione Peroxidase 1 was done on HepG2 cells. The cells were fixed, permeabilized and stained with Glutathione Peroxidase 1 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). 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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