The Peptide YY (PYY) BioAssay(TM) ELISA Kit is a sandwich ELISA for in vitro quantitative measurement of PYY in mouse serum, plasma and other biological fluids. Detection Detection Range: 15.63-1000pg/ml Sensitivity: 9.38pg/ml Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <10% Specificity: Recognizes mouse PYY. No significant crossreactivity or interference between mouse PYY and analogues was observed. Test Principle: This BioAssay(TM) ELISA kit uses Sandwich ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to mouse PYY. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for mouse PYY and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain mouse PYY, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450nm 2nm. The OD value is proportional to the concentration of mouse PYY. The concentration of mouse PYY in samples can be calculated by comparing the OD of the samples to the standard curve. Kit Components: *384210A: Microtiter Strips, 1x96 wells (8x12wells), Pre-coated, ready to use *384210B: Standard, 2x1vial 384210C: Standard Diluent, 1x20ml *384210D: Biotin Detection, 100X, 1x120ul *384210E: HRP Conjugate, 100X, 1x120ul 384210F: Biotin Detection Diluent, 1x14ml 384210G: HRP Diluent, 1x14ml 384210H: Substrate, 1x10ml 384210K: Stop Solution, 1x10ml 384210L: Wash Buffer, 25X, 1x30ml Precaution: The Stop Solution (384210K) included for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *384210A, *384210B, *384210D and *384210E at -20C. Store all the other components at 4C. Unused kit is stable for 6 months after receipt. 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 100ul standard or sample to each well. Incubate 90 minutes at 37C. 3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate. Incubate 15 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately
