Nori Porcine Cappase 8 ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of CASP8 in porcine. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for CASP8 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CASP8 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for CASP8 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of CASP8 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for caspase 8: CASP8
This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Porcine Caspase 8 ELISA Kit Summary
Alternative names for caspase 8: CASP8
Alternative names for porcine: pig, swine
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | A0A4X1V6R2 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 30 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Porcine Caspase 8 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
Caspase-8 (CASP8) is an apoptosis-related cysteine peptidase encoded by the CASP8 gene. CASP8 orthologs have been identified in nearly all mammals. Caspase-8 is a member of the caspase family of proteins, and has been shown to be an executioner protein of apoptosis. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes that undergo proteolytic processing by upstream caspases (caspase-8, -9) at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme in the form of a heterotetramer. The precursor of this caspase is cleaved by caspase 3, caspase 10, and caspase 9. It is activated upon cell death stimuli and induces apoptosis. Alternative splicing results in four transcript variants, encoding three distinct isoforms. Caspase 7 has been shown to interact with Caspase 8,[1] Survivin[2] and XIAP.[3][4][5] An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7.[2]
References
- Srinivasula SM, et al. (1996). Proc. Natl. Acad. Sci. U.S.A. 93 (25): 14486–91.
- Shin S, et al. (2001). Biochemistry. 40 (4): 1117–23.
- Roy N, et al. (1997). EMBO J. 16 (23): 6914–25.
- Deveraux QL, et al. (1997). Nature. 388 (6639): 300–4.
- Suzuki Y, et al. (2001). J. Biol. Chem. 276 (29): 27058–63.
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