This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti CASP5 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with CASP5 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of CASP5 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Specification
| Product Name | Human CASP5(Caspase-5) ELISA Kit |
| Alias | Caspase-5 ELISA Kit, CASP-5 ELISA Kit, ICE(rel)-III ELISA Kit, Protease ICH-3 ELISA Kit, Protease TY ELISA Kit, Caspase-5 subunit p20 ELISA Kit, Caspase-5 subunit p10 ELISA Kit, CASP5 ELISA Kit, ICH3 ELISA Kit |
| Sample Type | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples |
| Detection Method | Sandwich ELISA, Double Antibody |
| Detection Wavelength | OD450 |
| Reaction Duration | 4 hours |
| Storage | 2-8°C(Sealed), Don't cryopreserve. |
| Specificity | Specifically binds with CASP5 , no obvious cross reaction with other analogues. |
| UniProt ID | P51878 |
Standard Curve
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.). Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
| STD.(ng/ml) | OD-1 | OD-2 | Average |
| 0 | 0.071 | 0.073 | 0.072 |
| 78.125 | 0.147 | 0.151 | 0.149 |
| 156.25 | 0.225 | 0.231 | 0.228 |
| 312.5 | 0.493 | 0.507 | 0.5 |
| 625 | 0.799 | 0.823 | 0.811 |
| 1250 | 1.222 | 1.258 | 1.24 |
| 2500 | 1.769 | 1.821 | 1.795 |
| 5000 | 2.421 | 2.491 | 2.456 |
Recovery
Add a certain amount of CASP5 into the sample. Calculate the recovery by comparing the measured value with the expected amount of CASP5 in the sample.
| Sample Type | Recovery Range(%) | Average(%) |
| serum(n=10) | 84-100 | 95 |
| EDTA plasma(n=10) | 96-97 | 96 |
| Heparin plasma(n=10) | 83-99 | 89 |
Linearity
Dilute the sample with a certain amount of CASP5 at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 83-91% | 83-101% | 80-95% |
| EDTA plasma(n=10) | 84-103% | 84-97% | 83-93% |
| Heparin plasma(n=10) | 81-104% | 85-94% | 80-98% |
Precision(%)
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
| Item | Intra-assay Precision | Inter-assay Precision | ||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/ml) | 156.9 | 611.6 | 2443 | 160.7 | 633.6 | 2428 |
| Standard deviation | 8.38 | 31.38 | 127.04 | 8.16 | 31.81 | 128.68 |
| CV(%) | 5.34 | 5.13 | 5.2 | 5.08 | 5.02 | 5.3 |
Stability
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
| ELISA kit(n=5) | 37°C for 1 month | 2-8°C for 6 months |
| Average(%) | 80 | 95-100 |
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