| 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 sandwich ELISA quantifies human caspase-3 (gene CASP3) in cell culture supernatant sand serum. Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
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-Human CASP3 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human CASP3 Standard | 2 vials, 10 ng/tube |
| Human CASP3 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; 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.
Kit components are not sold separately.
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 |
| Intra-assay CV | 5.2–7.3% (3 samples, n = 16 each) |
| Inter-assay CV | 6.1–8.8% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.052; 31.2 = 0.0995; 62.5 = 0.1488; 125 = 0.2468; 250 = 0.4455; 500 = 0.724; 1000 = 1.232; 2000 = 1.994. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.081 | 0.202 | 0.29 | 0.418 | 0.672 | 0.998 | 1.642 | 2.633 |
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 Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- 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.
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