Oct 4 (2D5) Monoclonal Antibody

N2A cells were stained with Oct 4 (2D5) Monoclonal Antibody (bsm-52002R) at [1:200] incubated overnight at 4C, followed by secondary antibody incubation, DAPI staining of the nuclei and detection.

Western blot analysis of Oct4 on F9 cell lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/1,000 dilution.\r\n\r\nLysates\/proteins at 10 \u788cg\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 2 minutes;\r\n\r\n12% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) at 1\/1,000 dilution was used in 5% NFDM\/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.\r\n

Western blot analysis of Oct4 on different lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/500 dilution.\r\n\r\nLane 1: HES cell lysate\r\nLane 2: NCCIT cell lysate\r\n\r\nLysates\/proteins at 10 \u00b5g\/Lane.\r\n\r\nPredicted band size: 39 kDa\r\nObserved band size: 45 kDa\r\n\r\nExposure time: 1 minute;\r\n\r\n10% SDS-PAGE gel.\r\n\r\nProteins were transferred to a PVDF membrane and blocked with 5% NFDM\/TBST for 1 hour at room temperature. The primary antibody (bsm-52002R) 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.\r\n

Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-Oct4 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 (bsm-52002R, 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.

ICC staining of Oct4 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 (bsm-52002R, 1\/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor\u00ae488 Goat anti-Rabbit IgG was used as the secondary antibody at 1\/1,000 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemical analysis of paraffin-embedded human seminoma tissue tissue with Rabbit anti-Oct4 antibody (bsm-52002R) at 1\/4,000 dilution.\r\n\r\nThe section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (bsm-52002R) at 1\/4,000 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.