| Field | Specification |
|---|---|
| Target | |
| Alternative names | Caspase-8;CASP-8;3.4.22.61;Apoptotic cysteine protease;Apoptotic protease Mch-5;CAP4;FADD-homologous ICE/ced-3-like protease;FADD-like ICE;FLICE;ICE-like apoptotic protease 5;MORT1-associated ced-3 homolog;MACH;Caspase-8 subunit p18;Caspase-8 subunit p10;CASP8;MCH5; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: S217-D384 & L385-D479 |
| 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-8 (gene CASP8) in cell culture supernatantsand 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-8 is the initiator caspase of the extrinsic apoptosis pathway. Recruited through FADD to death receptors such as Fas and TNFR1, it becomes active and cleaves caspase-3, caspase-7 and BID. Caspase-8 also restrains necroptosis by cleaving RIPK1 and RIPK3, which makes it a central switch between apoptosis, necroptosis and inflammation.
Kit Components
| Anti-Human CASP8 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human CASP8 Standard | 2 vials, 10 ng/tube |
| Human CASP8 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 supernatantsand serum |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human CASP8 (Ser217–Asp384), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.4–6.4% (3 samples, n = 16 each) |
| Inter-assay CV | 5.4–8.2% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum samples measured about 120 pg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.019; 31.2 = 0.08; 62.5 = 0.131; 125 = 0.219; 250 = 0.437; 500 = 0.808; 1000 = 1.531; 2000 = 2.62. 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. | 0.019 | 0.08 | 0.131 | 0.219 | 0.437 | 0.808 | 1.531 | 2.62 |
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) | 89 | 257 | 911 | 90 | 251 | 854 |
| Standard deviation | 6.49 | 16.44 | 46.46 | 6.57 | 20.58 | 46.11 |
| CV (%) | 6.4% | 6.4% | 5.4% | 7.3% | 8.2% | 5.4% |
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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Sarah M. El-Demiry et al. (2022) Necroptosis modulation by cisplatin and sunitinib in hepatocellular carcinoma cell line. Life Sciences. 10.1016/j.lfs.2022.120594
Tang Dong et al. (2015) Apoptosis and anergy of T cell induced by pancreatic stellate cells-derived galectin-1 in pancreatic cancer. Tumor Biology. 10.1007/s13277-015-3233-5
