The Neuropeptide Y (NPY) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of NPY in bovine serum, plasma, tissue homogenates, cell lysates, cell culture supernates and and other biological fluids.
Detection Range:24.69-2000pg/ml
Sensitivity:9.35pg/ml
Intra-Assay CV:<10%
Inter-Assay CV:<12%
Test Principle:This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for NPY has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled NPY and unlabeled NPY (standards or samples) with the pre-coated antibody specific for NPY. 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 NPY in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of NPY in the sample.
Precaution:The Stop Solution (027111K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Storage and Stability:Store *027111A, *027111B, *027111D *027111E and *027111J at -20°C. After reconstitution of *027111J, store at 4°C and use within 5 days. 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 Summary:1. Prepare all reagents, samples and standards
2. Add 50ul standard or sample to each well, 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 absorbance at 450nm immediately.