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ELK Biotechnology

ELK9738

Rat KYN(Kynurenine) ELISA Kit

Rat KYN(Kynurenine) ELISA Kit

Sensitivity

Rat KYN(Kynurenine) ELISA Kit

Overview

Product name: Rat KYN(Kynurenine) ELISA Kit
Reactivity: Rat
Alternative Names: KYN; Kynurenine
Assay Type: Sandwich
Sensitivity: 3.5 pmol/ml
Standard: 500 pmol/ml
Detection Range: 7.82-500 pmol/ml
Sample Type: serum, plasma, tissue homogenates, saliva, cell lysates, follicular fluid and other biological fluids
Assay Length: 3.5h
Research Area: Metabolism
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 Rat KYN. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat KYN. 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 Rat KYN, 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 Rat KYN 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
500.00 2.354 2.267
250.00 1.647 1.560
125.00 1.088 1.001
62.50 0.821 0.734
31.25 0.567 0.480
15.63 0.336 0.249
7.82 0.252 0.165
0.00 0.087 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 KYN and the recovery rates were calculated by comparing the measured value to the expected amount of KYN in samples.
Matrix Recovery range Average
serum(n=5) 82-95% 88%
EDTA plasma(n=5) 90-103% 96%
Heparin plasma(n=5) 90-105% 97%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of KYN 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% 87-101% 89-99% 89-97%
EDTA plasma(n=5) 86-95% 81-93% 91-103% 85-99%
Heparin plasma(n=5) 82-96% 87-96% 89-98% 83-103%
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