EasyPro Series ELISA Kits
EasyPro Universal Biotin Quantitative Determination ELISA Kit
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Product No
FY-EU10029SSample Type
Serum, Plasma, Other biological fluids,Sample volume: 50μLAssay Time
45minPurchase details
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Product Details
Product Name in English:
EasyPro Universal Biotin Quantitative Determination ELISA Kit
Species:
Universal
Alternative Name:
Vitamin H, Coenzyme R, VB7, Vitamin B7, Biopeiderm
Assay Type:
Competitive
Standard:
500pg/mL
Sensitivity:
5.35 pg/mL
Detection Range:
7.81-500 pg/mL
Assay Time:
45min
Sample Type:
Serum, Plasma, Other biological fluids,Sample volume: 50μL
Research:
Immunology
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Biotin Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for Biotin are added to the micro ELISA plate wells. Biotin in samples (or standards) competes with a fixed amount of Biotin on the solid phase supporter for sites on the HRP linked detection antibody specific to Biotin. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The concentration of Biotin 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 |
|---|---|---|---|---|
| 500 | 0.168 | 0.17 | 0.169 | |
| 250 | 0.306 | 0.307 | 0.307 | |
| 125 | 0.525 | 0.528 | 0.527 | |
| 62.5 | 0.856 | 0.867 | 0.862 | |
| 31.25 | 1.277 | 1.294 | 1.286 | |
| 15.63 | 1.734 | 1.761 | 1.748 | |
| 7.82 | 2.043 | 2.069 | 2.056 | |
| 0 | 2.523 | 2.554 | 2.539 |
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) | 87-99 | 93 |
| EDTA plasma (n=8) | 80-95 | 86 |
| Cell culture media (n=8) | 95-107 | 101 |
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 (%) | 86-96 | 80-101 | 95-103 |
| Average (%) | 96 | 85 | 96 | |
| 1:4 | Range (%) | 85-97 | 87-96 | 87-98 |
| Average (%) | 96 | 91 | 91 | |
| 1:8 | Range (%) | 89-101 | 96-105 | 85-92 |
| Average (%) | 91 | 105 | 91 | |
| 1:16 | Range (%) | 85-97 | 95-103 | 80-96 |
| Average (%) | 89 | 96 | 80 |
Assay Procedures
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