Mini Samples ELISA Kit for Ghrelin (GHRL)

MTLRP; Growth Hormone-Releasing Peptide

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
  • Mini Samples ELISA Kit for Ghrelin (GHRL)Packages (Simulation)
  • Mini Samples ELISA Kit for Ghrelin (GHRL)Packages (Simulation)
  • Mini Samples ELISA Kit for Ghrelin (GHRL)Results demonstration
  • MEA991Mu.jpgTypical Standard Curve
  • CertificateISO9001: 2008, ISO13485: 2003 Registered

Specificity

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

Recovery

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

MatrixRecovery range (%)Average(%)
serum(n=5)90-10194
EDTA plasma(n=5)88-9591
heparin plasma(n=5)90-9895

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ghrelin (GHRL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ghrelin (GHRL) 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 Ghrelin (GHRL) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample1:21:41:81:16
serum(n=5)79-96%83-101%83-90%93-101%
EDTA plasma(n=5)91-105%80-93%87-99%93-101%
heparin plasma(n=5)78-92%98-105%80-97%87-94%

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

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×60µLAssay Diluent A1×6mL
Detection Reagent B1×60µLAssay Diluent B1×6mL
TMB Substrate1×3mLStop Solution1×3mL
Wash Buffer (30 × concentrate)1×10mLInstruction manual1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

Mini Samples ELISA Kit for Ghrelin (GHRL)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Ghrelin (GHRL) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Ghrelin (GHRL) and unlabeled Ghrelin (GHRL) (Standards or samples) with the pre-coated antibody specific to Ghrelin (GHRL). 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 Ghrelin (GHRL) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Ghrelin (GHRL) in the sample.

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