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 NOS2/iNOS antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with NOS2/iNOS 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 NOS2/iNOS in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Specification
| Product Name | Human NOS2/iNOS(Nitric oxide synthase, inducible) ELISA Kit |
| Alias | Nitric oxide synthase, inducible ELISA Kit, Hepatocyte NOS ELISA Kit, HEP-NOS ELISA Kit, Inducible NO synthase ELISA Kit, Inducible NOS, iNOS ELISA Kit, NOS type II ELISA Kit, Peptidyl-cysteine S-nitrosylase NOS2 ELISA Kit, NOS2 ELISA Kit, NOS2A ELISA Kit |
| UniProt ID | P35228 |
| 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 NOS2/iNOS , no obvious cross reaction with other analogues. |
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.(pg/ml) | OD-1 | OD-2 | Average |
| 0 | 0.155 | 0.159 | 0.157 |
| 78.125 | 0.211 | 0.217 | 0.214 |
| 156.25 | 0.348 | 0.358 | 0.353 |
| 312.5 | 0.463 | 0.477 | 0.47 |
| 625 | 0.719 | 0.739 | 0.729 |
| 1250 | 1.052 | 1.082 | 1.067 |
| 2500 | 1.563 | 1.609 | 1.586 |
| 5000 | 2.234 | 2.298 | 2.266 |
Recovery
Add a certain amount of NOS2/iNOS into the sample. Calculate the recovery by comparing the measured value with the expected amount of NOS2/iNOS in the sample.
| Sample Type | Recovery Range(%) | Average Recovery(%) |
| serum(n=10) | 86-100 | 93 |
| EDTA plasma(n=10) | 90-101 | 95 |
| Heparin plasma(n=10) | 95-102 | 99 |
Linearity
Dilute the sample with a certain amount of NOS2/iNOS at 1:2, 1:4 and 1:8 to get the recovery range.
| Sample Type | 1:2 | 1:4 | 1:8 |
| serum(n=10) | 95-105% | 94-104% | 89-95% |
| EDTA plasma(n=10) | 86-100% | 84-98% | 83-98% |
| Heparin plasma(n=10) | 87-93% | 82-99% | 81-100% |
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) | 152.6 | 593.5 | 2443 | 156.6 | 615.5 | 2427 |
| Standard deviation | 7.95 | 29.67 | 123.86 | 7.75 | 30.1 | 125.48 |
| CV(%) | 5.21 | 5 | 5.07 | 4.95 | 4.89 | 5.17 |
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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