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Human VD3(Vitamin D3) ELISA Kit
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ELK0811
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48 T $265.00
96 T $376.00
96 T*5 $1598.00
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OverviewPublications(1)

Product name: Human VD3(Vitamin D3) ELISA Kit
Reactivity: Human
Alternative Names: Cholecalciferol; Toxiferol; Activated 7-Dehydrocholesterol
Assay Type: Competitive Inhibition
Sensitivity: 2.07 ng/mL
Standard: 400 ng/mL
Detection Range: 6.25-400 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Metabolic pathway;Reproductive 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 Human VD3. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human VD3. 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 Human VD3 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
400.00 0.232
200.00 0.336
100.00 0.586
50.00 0.895
25.00 1.297
12.50 1.578
6.25 1.869
0.00 2.112

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 VD3 and the recovery rates were calculated by comparing the measured value to the expected amount of VD3 in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 89-103% 96%
Heparin plasma(n=5) 90-105% 97%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of VD3 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) 82-95% 89-102% 85-92% 96-104%
EDTA plasma(n=5) 98-106% 87-98% 83-95% 84-99%
Heparin plasma(n=5) 78-92% 83-96% 95-107% 87-102%