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☑ Cell treatment (CT)
Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (HA750159) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM staurosporine for 4 hours cell lysate
Lane 3: HeLa treated with 3μM staurosporine for 4 hours cell lysate
Lane 4: C6 cell lysate
Lane 5: C6 treated with 1μM staurosporine for 4 hours cell lysate
Lane 6: C6 treated with 3μM staurosporine for 4 hours cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 32/17 kDa
Observed band size: 32/17 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% 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 (HA750159) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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☑ Knockout (KO)
Western blot analysis of Caspase-3 with anti-Caspase-3 antibody (HA750159) at 1:2,000 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2: Caspase-3 knockout Hela whole cell lysate (10 µg).
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (HA750159, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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☑ Cell treatment (CT)
Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (HA750159) at 1/5,000 dilution.
Lane 1: U-87 MG cell lysate
Lane 2: MDA-MB-231 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: Jurkat treated with 1μM staurosporine for 3 hours cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 32/17 kDa
Observed band size: 32/17 kDa
Exposure time: 1 minute 2 seconds;
4-20% 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 (HA750159) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (HA750159) at 1/1,000 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 (HA750159) at 1/1,000 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 spleen tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (HA750159) at 1/1,000 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 (HA750159) at 1/1,000 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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