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Hyp(Hydroxyproline) ELISA Kit
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ELK7835
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48 T $265.00
96 T $376.00
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Overview

Product name: Hyp(Hydroxyproline) ELISA Kit
Reactivity: General
Alternative Names: (2S,4R)-4-Hydroxyproline; L-Hydroxyproline(C5H9O3N)
Assay Type: Competitive Inhibition
Sensitivity: 25.4 ng/mL
Standard: 5000 ng/mL
Detection Range: 78.13-5000 ng/mL
Sample type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay length: 2h
Research Area: Metabolic pathway;Nutrition metabolism;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Hyp. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Hyp. 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 450nm ± 10nm. The concentration of Hyp 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
5000.00 0.227
2500.00 0.396
1250.00 0.557
625.00 0.861
312.50 1.119
156.25 1.565
78.13 1.793
0.00 2.211

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 Hyp and the recovery rates were calculated by comparing the measured value to the expected amount of Hyp in samples.
Matrix Recovery range Average
serum(n=5) 87-99% 93%
EDTA plasma(n=5) 82-95% 88%
Heparin plasma(n=5) 90-105% 95%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Hyp 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% 92-105% 96-105% 95-103%
EDTA plasma(n=5) 95-104% 82-96% 92-105% 83-92%
Heparin plasma(n=5) 80-101% 87-101% 86-97% 88-95%