| The Colipase, Pancreatic (CLPS) BioAssay(TM) ELISA Ki is a sandwich enzyme immunoassay for in vitro quantitative measurement of CLPS in porcine serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Detection Range: 0.312-20ng/ml Sensitivity: 0.133ng/ml Precision: Intra-Assay: CV<10% Inter-Assay: CV<12% Assay Principle: The microplate provided in this kit has been pre-coated with an antibody specific to CLPS. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to CLPS. 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 CLPS, 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 CLPS in the samples is then determined by comparing the O.D. of the samples to the standard curve. Kit Components: Microtiter Strips, 1x96 wells, Pre-coated, ready to use Standard, 2x vial Detection Reagent A, 1*120ul Detection Reagent B, 1*120ul TMB Substrate, 1*9ml Wash Buffer (30X), 1*20ml Standard Diluent, 1*20ml Assay Diluent A, 1*12ml Assay Diluent B, 1*12ml Stop Solution, 1x6ml Precaution: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Assay Procedure Summary: 1. Prepare all reagents, samples and standards, 2. Add 50ul standard or sample to each well. 3. Add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate Solution. Incubate 10-20 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately. Storage and Stability: Store Microtiter Strips, Standard, Detection Reagent A, Detection Reagent B and Positive Control at -20C. After reconstitution of the positive control, store at 4C and use within 5 days. 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. |
