ELISA Kit for Corticotropin Releasing Hormone (CRH)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
- Product No.CEA835Hu
- Organism SpeciesHomo sapiens (Human)
- Brand OwnerCloud-Clone Corp.(USA)
- ManufacturerCloud-Clone Corp.Wuhan
- Test MethodCompetitive
- Assay Length2.5 hours
- Detection Range12.35-1000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 4.55pg/mL.
- Sample TypeSerum, plasma and other biological fluids
- DownloadInstruction Manual
- FOBUS$ 441US$ 630US$ 2835US$ 5355US$ 44100
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.
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(%)|
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
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.
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
|Pre-coated, ready to use 96-well strip plate||1||Plate sealer for 96 wells||4|
|Detection Reagent A||1||Assay Diluent A||1×12mL|
|Detection Reagent B||1×120µL||Assay Diluent B||1×12mL|
|Reagent Diluent||1×300µL||Stop Solution||1×6mL|
|TMB Substrate||1×9mL||Instruction manual||1|
|Wash Buffer (30 × concentrate)||1×20mL|
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 15-25 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Corticotropin Releasing Hormone (CRH) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Corticotropin Releasing Hormone (CRH) and unlabeled Corticotropin Releasing Hormone (CRH) (Standards or samples) with the pre-coated antibody specific to Corticotropin Releasing Hormone (CRH). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. 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.