active+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04]

☑ Cell treatment (CT)

Western blot analysis of active+pro Caspase-3 on different lysates with Rabbit anti-active+pro Caspase-3 antibody (ET1608-64) 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;

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 (ET1608-64) 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.

☑ Knockout (KO)

All lanes: Western blot analysis of Caspase-3 with anti-Caspase-3 antibody (ET1608-64) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2: Caspase-3 knockout Hela whole cell lysate (10 µg).

ET1608-64 was shown to specifically react with Caspase-3 in wild-type Hela cells. No band was observed when Caspase-3 knockout sample was tested. Wild-type and Caspase-3 knockout 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 (ET1608-64, 1/500) was used in 5% BSA 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.

☑ Cell treatment (CT)

Western blot analysis of active+pro Caspase-3 on different lysates with Rabbit anti-active+pro Caspase-3 antibody (ET1608-64) 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 (ET1608-64) 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.

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-active+pro Caspase-3 antibody (ET1608-64) 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 (ET1608-64) 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.

Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-active+pro Caspase-3 antibody (ET1608-64) 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 (ET1608-64) 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.

Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-active+pro Caspase-3 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 (ET1608-64, 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.

Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-active+pro Caspase-3 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 (ET1608-64, 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.