ELISA Kit for Corticotropin Releasing Hormone (CRH)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species Homo sapiens (Human)
Product No.CEA835Hu
Sample typeSerum, plasma and other biological fluids.
Format96-well strip plate
Assay length2.5 hours
Detection range12.35-1000pg/mL The standard curve concentrations used for the ELISA’s were 1000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL
SensitivityThe minimum detectable dose of this kit is typically less than 4.55pg/mL.
Specificity
This assay has high sensitivity and excellent specificity for detection of Corticotropin Releasing Hormone (CRH).
No significant cross-reactivity or interference between Corticotropin Releasing Hormone (CRH) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Corticotropin Releasing Hormone (CRH) and the recovery rates were calculated by comparing the measured value to the expected amount of Corticotropin Releasing Hormone (CRH) in samples.
Matrix Recovery range (%) Average(%)
serum(n=5)85-9389
EDTA plasma(n=5)91-10195
heparin plasma(n=5)93-10197
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Corticotropin Releasing Hormone (CRH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Corticotropin Releasing Hormone (CRH) 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 Corticotropin Releasing Hormone (CRH) 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)86-101%78-102%83-99%97-104%
EDTA plasma(n=5)83-101%80-105%90-98%80-93%
heparin plasma(n=5)78-92%94-101%85-99%97-105%
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 37oC;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test principle
This assay employs the competitive inhibition enzyme immunoassay technique. An antibody specific for Corticotropin Releasing Hormone (CRH) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between Horseradish Peroxidase (HRP) labeled Corticotropin Releasing Hormone (CRH) and unlabeled Corticotropin Releasing Hormone (CRH) (Standards or samples) with the pre-coated antibody specific for Corticotropin Releasing Hormone (CRH). After incubation the unbound conjugate is washed off. The amount of bound HRP conjugate is reverse proportional to the concentration of Corticotropin Releasing Hormone (CRH) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Corticotropin Releasing Hormone (CRH) in the sample.
Package and Components
Reagent Preparation
Results demonstration
Typical Standard Curve
Certificate
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Free trial
In case it is difficult to pre-evaluate experiment result because of specific test samples or other reasons, free trial can be offered.
If the budget is really limited or other special situation, you can apply the free products.
Guarantees
Unconditionally return policy within 72 hours.
For any complaint, we promise to reply within one work day, resolve the problem within three work days. Implementing initial consultation system in order to provide satisfactory service for all the customers. Customers can unconditionally to request a charge back or refund.
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