active+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04]
cat.: ET1608-64
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IP
Clonality: Monoclonal
Clone number: SU38-04
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 32/17 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human Caspase-3 aa 9-175 / 277.
Positive control: HeLa cell lysate, HeLa treated with 1μM staurosporine for 4 hours cell lysate, HeLa treated with 3μM staurosporine for 4 hours cell lysate, C6 cell lysate, C6 treated with 1μM staurosporine for 4 hours cell lysate, C6 treated with 3μM staurosporine for 4 hours cell lysate, U-87 MG cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, Jurkat cell lysate, Jurkat treated with 1μM staurosporine for 3 hours cell lysate, human lung carcinoma tissue, human spleen tissue, human liver tissue, human kidney tissue.
Subcellular location: Cytoplasm.
Recommended Dilutions:
  WB
  IHC-P
  IP

1:1,000-1:2,000
1:50-1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P42574 Human | P70677 Mouse | P55213 Rat
Alternative names: Caspase3 A830040C14Rik Apopain CASP 3 CASP-3 CASP3 CASP3_HUMAN Casp3a Caspase 3 Caspase 3, apoptosis-related cysteine peptidase Caspase 3, apoptosis-related cysteine protease Caspase 3, apoptosis-related cysteine protease a Caspase-3 subunit p12 Caspase3 CC3 CPP 32 CPP-32 CPP32 CPP32B Cysteine protease CPP32 EC 3.4.22.56 ICE3 LICE mldy OTTHUMP00000165052 OTTHUMP00000165053 OTTHUMP00000165054 PARP cleavage protease Procaspase3 Protein Yama SCA 1 SCA-1 SCA1 SREBP cleavage activity 1 Yama Yama protein
Images
ET1608-64_1.jpg Fig1: 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.
ET1608-64_2.jpg Fig2: 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.
ET1608-64_3.jpg Fig3: 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.
ET1608-64_4.jpg Fig4: 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.
ET1608-64_5.jpg Fig5: 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.
ET1608-64_6.jpg Fig6: 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.
ET1608-64_7.jpg Fig7: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.