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Alternative Names: NOS2A; INOS; HEP-NOS; I-NOS; Hepatocytes Oxide Synthase; Peptidyl-cysteine S-nitrosylase NOS2; NOS2; Nitric Oxide Synthase 2, Inducible
Catalogue No. 2643ELK
Size 96T
Reactivity Human
Range 0.16-10 ng/mL
Sensitivity 0.054 ng/mL

Description

Alternative Names: NOS2A; INOS; HEP-NOS; I-NOS; Hepatocytes Oxide Synthase; Peptidyl-cysteine S-nitrosylase NOS2; NOS2; Nitric Oxide Synthase 2, Inducible
Catalogue No. 2643ELK
Size 96T
Reactivity Human
Range 0.16-10 ng/mL
Sensitivity 0.054 ng/mL
Assay Type Sandwich
Sample Type serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length 3.5h
Research Area Enzyme & Kinase;Metabolic pathway;Tumor immunity;Cardiovascular biology;
Test principle             The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human NOS2/iNOS. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human NOS2/iNOS. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human NOS2/iNOS, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human NOS2/iNOS in the samples is then determined by comparing the OD of the samples to the standard curve.
Standard Curve
Concentration (ng/mL) OD Corrected OD
10.00 1.984 1.899
5.00 1.586 1.501
2.50 1.174 1.089
1.25 0.797 0.712
0.63 0.529 0.444
0.32 0.347 0.262
0.16 0.187 0.102
0.00 0.085 0.000

Precision Intra-assay Precision (Precision within an assay):CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays):CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery
Matrices listed below were spiked with certain level of recombinant NOS2/iNOS and the recovery rates were calculated by comparing the measured value to the expected amount of NOS2/iNOS in samples.
Matrix Recovery range Average
serum(n=5) 95-107% 101%
EDTA plasma(n=5) 85-95% 90%
Heparin plasma(n=5) 79-93% 86%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NOS2/iNOS and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 87-96% 86-98% 91-103% 87-90%
EDTA plasma(n=5) 81-94% 85-97% 93-102% 81-95%
Heparin plasma(n=5) 83-98% 92-103% 86-97% 93-101%
Note For Research Use Only

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