Acylated Ghrelin (A-GHR) BioAssay™ ELISA Kit (Human)

Kits and Assays / Kits-ELISA, BioAssay™

4°C/-20°C

The Acylated Ghrelin (A-GHR) BioAssay™ ELISA Kit (Human) is a quantitative sandwich assay for the detection of A-GHR in human serum, plasma and other biological fluids. Cell lysates and tissue homogenates, though not tested, may potentially be used as samples.

Detection Range:
15.625-1000pg/ml

Sensitivity:
<9.375pg/ml

Precision:
Inter-Assay CV: <10%
Intra-Assay CV: <8%

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to A-GHR. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to A-GHR. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain A-GHR, biotin-conjugated antibody and enzyme-conjugated streptavidin complex will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of A-GHR in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
*353564A: Microtiter Strips, 8x12 wells.
*353564B: Standard, 2 vials
353564C: Sample/Standard Dilution Buffer, 1x20ml
353564D: Antibody-Biotin (Concentrated), 1x120ul
353564E: Antibody Dilution Buffer, 1x10ml
353564F: Streptavidin-HRP Conjugate (SABC), 1x120ul
353564G: SABC Dilution Buffer, 1x10ml
353564H: TMB Substrate, 1x10ml
353564J: Stop Solution, 1x10ml
353564K: Wash Buffer (25X), 1x30ml

Storage and Stability:
Store unopened *353564A at 4ºC; store at -20ºC once opened. Store unopened *353564B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.

Assay Summary:
1. Wash plate 2 times before adding standards, samples and control (zero) to wells.
2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C.
3. Discard the liquid and wash plate 2 times.
3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C
4. Discard the liquid and wash plate 3 times.
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C
6. Discard the liquid and wash plate 5 times.
7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results.