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25-OH-D(25 Hydroxy Vitamin D) ELISA Kit
ELK9387
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48 T $265.00
96 T $376.00
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Overview

Product name: 25-OH-D(25 Hydroxy Vitamin D) ELISA Kit
Reactivity: General
Alternative Names: 25 Hydroxy Vitamin D;
Assay Type: Competitive Inhibition
Sensitivity: 0.91 ng/mL
Standard: 200 ng/mL
Detection Range: 3.13-200 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Reproductive science;Genetic science;Nutrition metabolism;Bone metabolism;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 25-OH-D protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 25-OH-D. 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 25-OH-D 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
200.00 0.199
100.00 0.357
50.00 0.587
25.00 0.823
12.50 1.253
6.25 1.596
3.13 1.871
0.00 2.233

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 25-OH-D and the recovery rates were calculated by comparing the measured value to the expected amount of 25-OH-D in samples.
Matrix Recovery range Average
serum(n=5) 82-94% 88%
EDTA plasma(n=5) 93-107% 100%
Heparin plasma(n=5) 95-107% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 25-OH-D 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) 95-102% 98-106% 87-98% 83-95%
EDTA plasma(n=5) 84-99% 83-101% 95-104% 85-92%
Heparin plasma(n=5) 97-103% 87-98% 95-102% 81-103%