VALIDATION IMAGES
Lane 1: F9; Lane 2: NCCIT; Lane 3: hES lysates probed with Oct 4 (2D5) Monoclonal Antibody (bsm-52002R) at 1:1000 overnight at 4˚C. Followed by a conjugated secondary antibody.
Western blot analysis of Oct4 on F9 cell lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1/1,000 dilution.
Lysates/proteins at 10 碌g/Lane.
Predicted band size: 39 kDa
Observed band size: 45 kDa
Exposure time: 2 minutes;
12% 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 (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.
Western blot analysis of Oct4 on different lysates with Rabbit anti-Oct4 antibody (bsm-52002R) at 1/500 dilution.
Lane 1: HES cell lysate
Lane 2: NCCIT cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 39 kDa
Observed band size: 45 kDa
Exposure time: 1 minute;
10% 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 (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.
Immunohistochemical analysis of paraffin-embedded human seminoma tissue tissue with Rabbit anti-Oct4 antibody (bsm-52002R) at 1/4,000 dilution.
The 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.