Human Active Caspase-3 Quantikine ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # KM300
This product is in process of discontinuation.
Discontinued Product
KM300 has been discontinued.
View all Caspase-3 products.
Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Extracts (100 µL)
Sensitivity
0.1 ng/mL
Assay Range
0.31-20 ng/mL (Cell Extracts)
Product Summary for Human Active Caspase-3 Quantikine ELISA Kit
The Quantikine Human active Caspase-3 Immunoassay is a 3.5 hour solid phase ELISA designed to measure human active caspase-3 in cell extracts. It contains biotinylated-inhibitor derivatized recombinant caspase-3 and a monoclonal antibody raised against caspase-3. Results obtained using natural active caspase-3 covalently bound to biotin-ZVKD-fmk inhibitor in apoptotic cell extracts showed linear curves that were parallel to the Quantikine kit standard. These results indicate that this kit can be used to determine relative mass values for natural active caspase-3.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Biotin-ZVKD-fmk modified natural and biotin-D-fmk modified recombinant active Caspase-3. This assay also detects mouse active Caspase-3.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Extracts
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 1.06 | 3.09 | 7.70 | 1.06 | 3.27 | 8.25 |
| Standard Deviation | 0.05 | 0.16 | 0.20 | 0.07 | 0.19 | 0.50 |
| CV% | 4.7 | 5.2 | 2.6 | 6.6 | 5.8 | 6.1 |
Recovery for Human Active Caspase-3 Quantikine ELISA Kit
The recovery of active caspase-3 spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Calibrator Diluent RD5-20 (1X) (n=4) | 99 | 98-100 |
| Extraction Buffer (1X) (n=4) | 110 | 104-114 |
| Extracts from Jurkat Cells Treated w/Staurosporine (n=4) | 92 | 87-95 |
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of active caspase-3 in each matrix were diluted with Calibrator Diluent and then assayed. Results from typical sample dilutions are
Scientific Data Images for Human Active Caspase-3 Quantikine ELISA Kit
Human Active Caspase-3 ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Caspase-3
Caspases are a family of cytosolic aspartate-specific cysteine proteases involved in the initiation and execution of apoptosis. They are expressed as latent zymogens and are activated by an autoproteolytic mechanism or by processing by other proteases (frequently other caspases). Human caspases can be subdivided into three functional groups: cytokine activation (caspase-1, -4, -5, and -13), apoptosis initiation (caspase-2, -8, -9, -and -10), and apoptosis execution (caspase-3, -6, and -7).
Caspases are regulated by a variety of stimili, including APAF1, CFLAR/FLIP, NOL3/ARC, and members of the inhibitor of apoptosis (IAP) family such as BIRC1/NAIP, BIRC2/cIAP-1, BIRC3/cIAP-2, BIRC4/XIAP, BIRC5/Survivin, and BIRC7/Livin. IAP activity is modulated by DIABLO/SMAC or PRSS25/HTRA2/Omi. Cell-permeable and irreversible peptide inhibitors are also available for different caspases.
Alternate Names
Apopain, CASP3, Caspase3, CPP32, LICE-1, YAMA
Gene Symbol
CASP3
Additional Caspase-3 Products
Product Documents for Human Active Caspase-3 Quantikine ELISA Kit
Product Specific Notices for Human Active Caspase-3 Quantikine ELISA Kit
For research use only
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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.