Universal ELISA Kits
Hyp(Hydroxyproline) ELISA Kit
Product No
FY-EU1199Sample Type
serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluidsAssay Time
1.5HPurchase details
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DataSheet 
Product Details
Product Name in English:
Hyp(Hydroxyproline) ELISA Kit
Species:
Universal
Alternative Name:
(2S,4R)-4-Hydroxyproline; L-Hydroxyproline(C5H9O3N)
Assay Type:
Competitive
Standard:
5000 ng/mL
Sensitivity:
26.3 ng/mL
Detection Range:
78.13-5000 ng/mL
Assay Time:
1.5H
Sample Type:
serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Research:
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 protein. 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 stop 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.
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 |
|---|---|---|---|---|
| 5000 | 0.169 | 0.17 | 0.17 | |
| 2500 | 0.307 | 0.309 | 0.308 | |
| 1250 | 0.526 | 0.531 | 0.529 | |
| 625 | 0.857 | 0.859 | 0.858 | |
| 312.5 | 1.278 | 1.284 | 1.281 | |
| 156.25 | 1.735 | 1.752 | 1.744 | |
| 78.13 | 2.044 | 2.045 | 2.045 | |
| 0 | 2.524 | 2.528 | 2.526 |
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) | 80-95 | 87 |
| EDTA plasma (n=8) | 86-99 | 92 |
| Cell culture media (n=8) | 80-95 | 87 |
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 (%) | 85-92 | 86-93 | 90-101 |
| Average (%) | 91 | 88 | 94 | |
| 1:4 | Range (%) | 80-96 | 86-93 | 92-106 |
| Average (%) | 91 | 91 | 104 | |
| 1:8 | Range (%) | 89-100 | 93-101 | 92-101 |
| Average (%) | 99 | 94 | 94 | |
| 1:16 | Range (%) | 85-96 | 86-97 | 80-95 |
| Average (%) | 91 | 96 | 90 |
Assay Procedures
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