ELISA Kit for Cortisol (Cor)

Hydrocortisone

Specificity

This assay has high sensitivity and excellent specificity for detection of Cortisol (Cor).
No significant cross-reactivity or interference between Cortisol (Cor) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Cortisol (Cor) and the recovery rates were calculated by comparing the measured value to the expected amount of Cortisol (Cor) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 94
EDTA plasma(n=5) 81-93 86
heparin plasma(n=5) 88-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cortisol (Cor) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cortisol (Cor) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cortisol (Cor) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 91-104% 98-105% 96-103% 89-97%
EDTA plasma(n=5) 80-101% 93-105% 83-101% 78-96%
heparin plasma(n=5) 89-103% 95-105% 96-104% 89-98%

Stability

The stability of 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.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Research in Veterinary Science Use of infrared thermography to assess the influence of high environmental temperature on rabbits ScienceDirect: S0034528813001380
British Journal of Nutrition Identification of sociodemographic and clinical factors associated with the levels of human b-defensin-1 and human b-defensin-2 in the human milk of Han Chinese Pubmed:24124699
Theriogenology Daily exposure to summer circadian cycles affects spermatogenesis, but not fertility in an in vivo rabbit model Pubmed:25442388
Animal Production Science Effect of high temperatures on breeding rabbit behaviour Publish:Source
JBCD Assessment of cortisol as salivary psychological stress marker in relation to temporomandibular disorders among a sample of dental students Index.Php: Jbcd
International Journal of Immunopathology & Pharmacology Sea buckthorn (Hippophae rhamnoides L.) oil protects against chronic stress-induced inhibitory function of natural killer cells in rats PubMed: 26684638
IET Micro & Nano Letters Surface plasmon resonance immunoassay for cortisol determination with a self-assembling denaturalised bovine serum albumin layer on surface plasmon resonance chip xpls:7393900
Optoelectronics Letters Fabrication and characterization of SPR chips with the modified bovine serum albumin article:10.1007
Journal of Dairy Science Effect of thermal stress on physiological, hormonal and haematological parameters in Tharparkar and Karan Fries calves publication:306322503
Acta Veterinaria Hungarica Relationship between the indexes of insulin resistance and metabolic status in dairy cows during early lactation acve-2017-0006
BMC Veterinary Research Hypothalamus-pituitary-adrenal axis involves in anti-viral ability through regulation of immune response in piglets infected by highly pathogenic porcine … Pubmed:29540178
Indian Journal of Animal Sciences Endurance exercise causes adverse changes in some hematological and physio-biochemical indices in ponies under high altitude stress condition :
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