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ELK Biotechnology

ELK9594

Human A-GHRL(AcylatedGhrelin) ELISA Kit

Human A-GHRL(AcylatedGhrelin) ELISA Kit

Sensitivity

Human A-GHRL(AcylatedGhrelin) ELISA Kit

Overview

Product name: Human A-GHRL(AcylatedGhrelin) ELISA Kit
Reactivity: Human
Alternative Names: A-GHRL
Assay Type: Sandwich
Sensitivity: 9.38 pg/mL
Standard: 1000 pg/mL
Detection Range: 15.63-1000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Cardiovascular;Signal Transduction;Cardiovascular;Metabolism;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human A-GHRL. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human A-GHRL. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human A-GHRL, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human A-GHRL in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
1000.00 1.946 1.861
500.00 1.572 1.487
250.00 1.282 1.197
125.00 0.952 0.867
62.50 0.518 0.433
31.25 0.357 0.272
15.63 0.194 0.109
0.00 0.085 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant A-GHRL and the recovery rates were calculated by comparing the measured value to the expected amount of A-GHRL in samples.
Matrix Recovery range Average
serum(n=5) 84-96% 90%
EDTA plasma(n=5) 87-95% 91%
Heparin plasma(n=5) 95-107% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of A-GHRL and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 85-97% 95-107% 92-103% 81-99%
EDTA plasma(n=5) 80-96% 93-105% 83-97% 95-104%
Heparin plasma(n=5) 82-93% 81-96% 79-91% 83-94%
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