ELISA Kit for Caspase 9 (CASP9)

APAF3; ICE-LAP6; MCH6; Apoptosis-Related Cysteine Peptidase; Cysteinyl Aspartate Specific Proteinases 9; Apoptotic protease-activating factor 3; ICE-like apoptotic protease 6

Specificity

This assay has high sensitivity and excellent specificity for detection of Caspase 9 (CASP9).
No significant cross-reactivity or interference between Caspase 9 (CASP9) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Caspase 9 (CASP9) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Caspase 9 (CASP9) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Reference
Reprod Sci. Role of mitochondria in ciprofloxacin-induced apoptosis in murine sperm cells. Pubmed: 23420823
Molecular Neurobiology Ursodeoxycholic Acid Ameliorates Apoptotic Cascade in the Rotenone Model of Parkinson’s Disease: Modulation of Mitochondrial Perturbations Pubmed:25502462
BMC Cancer Saikosaponin d induces cell death through caspase-3-dependent, caspase-3-independent andmitochondrial pathways in mammalian hepatic stellate cells. pubmed:27461108
Exp Lung Res Stretch-induced apoptosis in rat alveolar epithelial cells is mediated by the intrinsic mitochondrial pathway. pubmed:28394655
The American Journal of Chinese Medicine Saikosaponin a Induces Apoptosis through Mitochondria-Dependent Pathway in Hepatic Stellate Cells pubmed:28231747
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