Bovine Cortisol (COR) ELISA Kit
Cat# AE59031BO
Size : 96Tests
Brand : Abebio
Reagent Preparation
Results demonstration
Assay Procedure Summary
Certificate
Product Details
| Species Reactivity | Bovine (Bos taurus; Cattle) |
| UniProt | N/A |
| Abbreviation | COR |
| Alternative Names | Hydrocortisone |
| Range | 312.5 - 5000 pg/mL |
| Sensitivity | 156 pg/mL |
| Sample Type | Serum, Plasma, Other biological fluids |
| Detection Method | Sandwich |
| Analysis Method | Quantitive |
| Assay Duration | 1-4.5h |
| Sample Volume | 1-200 μL |
| Detection Wavelengt | 450 nm |
Test principle
This assay employs the competitive inhibition enzyme immunoassay technique. An antibody specific for COR has been pre-coated onto a microplate. Standards or samples and a biotin-conjugated antibody specific for COR are pipetted into the wells. A competitive inhibition reaction is launched between COR (Standards or samples) and biotin-conjugated COR with the pre-coated antibody specific for COR. After washing, streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of COR bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview
Cortisol is a steroid hormone, more specifically a glucocorticoid, which is produced by the zona fasciculata of the adrenal cortex. It is released in response to stress and a low level of blood glucose.
Its functions are to increase blood sugar through gluconeogenesis, to suppress the immune system, and to aid the metabolism of fat, protein, and carbohydrate. It also decreases bone formation.
Hydrocortisone (INN, USAN, BAN) is a name for cortisol when it is used as a medication. Hydrocortisone is used to treat people who lack adequate naturally generated cortisol. It is on the World Health Organization's List of Essential Medicines needed in a basic health system.
Components
Reagents | Quantity | Reagents | Quantity |
Assay plate (96 Wells) | 1 | Instruction manual | 1 |
Standard (lyophilized) | 2 | Sample Diluent | 1 x 20 mL |
Biotin-Conjugate (concentrate 100 x) | 1 x 120 μL | Biotin-Conjugate Diluent | 1 x 12 mL |
Streptavidin-HRP (concentrate 100 x) | 1 x 120 μL | Streptavidin-HRP Diluent | 1 x 12 mL |
Wash Buffer (concentrate 25 x) | 1 x 20 mL | Substrate Solution | 1 x 10 mL |
Stop Solution | 1 x 6 mL | Adhesive Films | 4 |
Specificity
This assay has high sensitivity and excellent specificity for detection of Bovine COR. No significant cross-reactivity or interference between Bovine COR and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Bovine COR and the recovery rates were calculated by comparing the measured value to the expected amount of Bovine COR in samples.
Sample Type | Number | Recovery range (%) | Average(%) |
Serum | 10 | 90-101 | 96 |
EDTA plasma | 10 | 89-97 | 93 |
Heparin plasma | 10 | 91-99 | 95 |
Precision
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Bovine COR and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample Type | 1：2 | 1：4 | 1：8 | 1：16 |
Serum | 78-89% | 81-99% | 92-103% | 95-105% |
EDTA plasma | 91-101% | 90-98% | 93-101% | 91-98% |
Heparin plasma | 92-103% | 93-102% | 92-99% | 91-101% |
Stability
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
Sample collection and storage
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Kits storage instructions
Store at 2-8°C. Please refer to Instruction Manual.
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