The Mouse Ghrelin (GHRL) ELISA Kit is a competitive inhibition immunoassay for in vitro quantitative measurement of mouse GHRL in serum, plasma, tissue homogenates and other biological fluids.
Detection Range:123.5-10,000pg/ml
Sensitivity:42.5pg/ml
Test Principle:This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for mouse GHRL has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled mouse GHRL and unlabeled mouse GHRL (standards or samples) with the pre-coated antibody specific for mouse 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 inversely proportional to the concentration of GHRL in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of GHRL in the sample.
Kit Components:*025268A: Microtiter Plate, 96-well strip plate. Pre-coated, ready to use*025268B: Standard, 2x1vial 025268C: Standard Diluent, 1x20ml*025268D: Detection Reagent A (green), 1x120ul *025268E: Detection Reagent B (red), 1x120ul 025268F: Assay Diluent A (2x), 1x6ml025268G: Assay Diluent B (2x), 1x6ml025268H: TMB Substrate, 1x9ml025268K: Stop Solution, 1x6ml025268L: Wash Buffer, 30X, 1x20ml
Storage and Stability:Store *025268A, *025268B, *025268D and *025268E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Assay Procedure Summary:1. Prepare all reagents, samples and standards. 2. Add 50ul standard or sample to each well, and then add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C. 3. Aspirate and wash 3 times. 4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C. 5. Aspirate and wash 5 times. 6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37°C. 7. Add 50ul Stop Solution. Read at 450 nm immediately
