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Alternative Names: ECNOS; NOS III; E-NOS; C-NOS; ENOS; CNOS; Constitutive NOS
Catalogue No. 1500ELK
Size 96T
Reactivity Human
Range 0.16-10 ng/mL
Sensitivity 0.059 ng/mL

Description

Alternative Names: ECNOS; NOS III; E-NOS; C-NOS; ENOS; CNOS; Constitutive NOS
Catalogue No. 1500ELK
Size 96T
Reactivity Human
Range 0.16-10 ng/mL
Sensitivity 0.059 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;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 NOS3. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human NOS3. 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 NOS3, 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 NOS3 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 2.119 2.022
5.00 1.606 1.509
2.50 1.138 1.041
1.25 0.898 0.801
0.63 0.534 0.437
0.32 0.388 0.291
0.16 0.215 0.118
0.00 0.097 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 NOS3 and the recovery rates were calculated by comparing the measured value to the expected amount of NOS3 in samples.

Matrix Recovery range Average
serum(n=5) 82-97% 89%
EDTA plasma(n=5) 80-95% 87%
Heparin plasma(n=5) 86-99% 92%
Linearity The linearity of the kit was assayed by testing samples spiked with appropriate concentration of NOS3 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) 95-104% 93-102% 79-94% 87-98%
EDTA plasma(n=5) 88-101% 90-98% 92-106% 85-97%
Heparin plasma(n=5) 89-97% 85-97% 87-96% 85-96%
Note For Research Use Only

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