| Field | Specification |
|---|---|
| Target | |
| Alternative names | Caspase-3;CASP-3;3.4.22.56;Apopain;Cysteine protease CPP32;CPP-32;Protein Yama;SREBP cleavage activity 1;SCA-1;Caspase-3 subunit p17;Caspase-3 subunit p12;CASP3;CPP32; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S29-D175 & A183-H277 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This single-incubation sandwich ELISA quantifies human caspase-3 (gene CASP3) in cell culture supernatant sand serum. The strips are coated with an anti-tag antibody. Samples and standards are added to the wells together with the kit’s antibody cocktail (Antibody A and Antibody B); the resulting antibody–target complex is held on the plate through the tag. After washing, TMB substrate is added and the HRP label on the detection antibody converts it to a colored product in proportion to the bound target. The reaction is stopped and the plate is read at 450 nm. The supplier markets the Quick format as a 90-minute assay.
Caspase-3 is the major executioner caspase of apoptosis. Activated by initiator caspases such as caspase-8 and caspase-9, it cleaves hundreds of cellular proteins, including PARP1, leading to the typical features of apoptotic cell death. Cleaved (active) caspase-3 is one of the most widely used apoptosis markers in Western blot and immunohistochemistry.
Kit Components
| Anti-tag Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human CASP3 Standard | 2 × 10 ng/tube |
| Human CASP3 Antibody A | 3 mL |
| Human CASP3 Antibody B | 3 mL |
| Sample Diluent | 15 mL |
| TBS-T Wash Buffer (25x) | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Adhesive Plate Sealers | 2 |
Also required (not supplied): microplate reader for 450 nm; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution; horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3 mm.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatant sand serum |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human CASP3 (Ser29–Asp175), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
Storage & Handling
Store at 4 °C for up to 6 months. Ships on gel ice and tolerates up to 3 days at room temperature; refrigerate on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.001 | 0.041 | 0.084 | 0.168 | 0.262 | 0.476 | 0.984 | 1.886 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 63 | 211 | 945 | 69 | 203 | 995 |
| Standard deviation | 3.27 | 15.4 | 55.75 | 4.2 | 17.86 | 62.68 |
| CV (%) | 5.2% | 7.3% | 5.9% | 6.1% | 8.8% | 6.3% |
Kit components
Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x)|1|12ml Color Developing Reagent (TMB)|1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|PieceMaterials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Automated plate washer (optional)
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
- Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.
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