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DHVD3(1,25-Dihydroxyvitamin D3) ELISA Kit
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Product No

FY-EU1116

Sample Type

Serum, Plasma and Other biological fluids

Assay Time

1.5H

Price($)

280 400

Size

48T 96T

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Product Details
Product Name in English:
DHVD3(1,25-Dihydroxyvitamin D3) ELISA Kit
Species:
Universal
Alternative Name:
DH-VD3; Calcitriol; Rocaltrol; Calcijex; 1,25-(OH)2D3; 1,25-Dihydroxycholecalciferol; 1,25-Dihydroxy Vitamin D3
Assay Type:
Competitive
Standard:
10000 pg/mL
Sensitivity:
49.5 pg/mL
Detection Range:
156.25-10000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Metabolic pathway;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 DHVD3 protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to DHVD3. 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 stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of DHVD3 in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Concentration OD1 OD2 Average OD Corrected
10000 0.261 0.263 0.262
5000 0.361 0.364 0.363
2500 0.555 0.56 0.558
1250 0.816 0.824 0.82
625 1.111 1.121 1.116
312.5 1.506 1.513 1.51
156.25 1.82 1.824 1.822
0 2.484 2.49 2.487
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 80-91 86-97 92-105
Average (%) 81 97 93
1:4 Range (%) 89-103 86-97 87-98
Average (%) 90 88 95
1:8 Range (%) 82-95 90-98 85-92
Average (%) 92 97 91
1:16 Range (%) 86-99 80-93 80-96
Average (%) 94 87 95
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
Item Intra-assay Precision Intra-assay Precision
Sample 3 3
Replicate 9 18
CV(%) 5 8
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 90-105 97
EDTA plasma (n=8) 80-95 87
Cell culture media (n=8) 83-95 89
Assay Procedures
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