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Alternative Names: Citrulline
Catalogue No. 0856ELK
Size 96T
Reactivity Human
Range 1.57-100 ng/mL
Sensitivity 0.51 ng/mL

Description

Alternative Names: Citrulline
Catalogue No. 0856ELK
Size 96T
Reactivity Human
Range 1.57-100 ng/mL
Sensitivity 0.51 ng/mL
Assay Type Competitive Inhibition
Sample Type serum, plasma, urine and other biological fluids
Assay Length 2h
Research Area Metabolism
Test principle This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Citrulline(Cit) protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Citrulline(Cit). Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 10 nm. The concentration of Citrulline(Cit) 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
100.00 0.189
50.00 0.386
25.00 0.583
12.50 0.857
6.25 1.152
3.13 1.511
1.57 1.762
0.00 2.115

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 Cit and the recovery rates were calculated by comparing the measured value to the expected amount of Cit in samples.

Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 81-95% 88%
Linearity The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cit 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) 85-94% 97-106% 92-101% 82-90%
EDTA plasma(n=5) 85-92% 97-103% 87-98% 95-102%
Heparin plasma(n=5) 85-104% 79-92% 83-95% 86-97%
Note For Research Use Only

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