EasyPro Series ELISA Kits
EasyPro Rat OXB(Orexin B) ELISA Kit
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Product No
FY-ER2130SSample Type
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μLAssay Time
45minPurchase details
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Product Details
Product Name in English:
EasyPro Rat OXB(Orexin B) ELISA Kit
Species:
Rat
Alternative Name:
Hypocretin-2
Assay Type:
Competitive
Standard:
10ng/mL
Sensitivity:
0.1 ng/mL
Detection Range:
0.16-10ng/mL
Assay Time:
45min
Sample Type:
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μL
Research:
Signal transduction;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with OXB Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for OXB are added to the micro ELISA plate wells. OXB in samples (or standards) competes with a fixed amount of OXB on the solid phase supporter for sites on the HRP linked detection antibody specific to OXB. 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 OXB 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 |
|---|---|---|---|---|
| 10 | 0.294 | 0.299 | 0.297 | |
| 5 | 0.394 | 0.4 | 0.397 | |
| 2.5 | 0.588 | 0.592 | 0.59 | |
| 1.25 | 0.849 | 0.855 | 0.852 | |
| 0.63 | 1.144 | 1.149 | 1.147 | |
| 0.32 | 1.539 | 1.56 | 1.55 | |
| 0.16 | 1.853 | 1.862 | 1.858 | |
| 0 | 2.517 | 2.566 | 2.542 |
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 (%) | 84-99 | 85-100 | 86-93 |
| Average (%) | 92 | 97 | 89 | |
| 1:4 | Range (%) | 83-95 | 84-98 | 91-96 |
| Average (%) | 87 | 86 | 95 | |
| 1:8 | Range (%) | 83-95 | 97-101 | 87-98 |
| Average (%) | 89 | 99 | 96 | |
| 1:16 | Range (%) | 96-101 | 88-96 | 95-102 |
| Average (%) | 100 | 92 | 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) | 89-103 | 96 |
| EDTA plasma (n=8) | 83-97 | 90 |
| Cell culture media (n=8) | 86-99 | 92 |
Assay Procedures
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