Myeloperoxidase (MPO) BioAssay(TM) ELISA Kit (Bovine)

Product Overview

• Article Name: Myeloperoxidase (MPO) BioAssay(TM) ELISA Kit (Bovine)
• Biozol Catalog Number: USB-026939
• Supplier Catalog Number: 026939
• Alternative Catalog Number: USB-026939-96
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

The Bovine Myeloperoxidase (MPO) ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of MPO in bovine serum, plasma, cell lysates and other biological fluids. Detection Range: 0.312-20ng/ml Sensitivity: <0.135ng/ml Precision: Intra-Assay: CV<10% Inter-Assay: CV<12% Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to MPO. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to MPO. 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 MPO, biotin-conjugated antibody and enzyme-conjugated Avidin 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 MPO in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *026939A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *026939B: Standard, 2x1vial 026939C: Standard Diluent, 1x20ml *026939D: Detection Reagent A, 1x120ul *026939E: Detection Reagent B, 1x120ul 026939F: Assay Diluent A, 1x12ml 026939G: Assay Diluent B, 1x12ml 026939H: TMB Substrate, 1x9ml 026939K: Stop Solution, 1x6ml 026939L: Wash Buffer, 30x, 1x20ml Precaution: The Stop Solution (026939K) 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 *026939A, *026939B, *026939D and *026939E at -20C. Store all the other components at 4C. 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. Materials Required But Not Supplied: 1. Microplate reader with 450 10nm filter. 2. Precision single or multi-channel pipettes and disposable tips. 3. Eppendorf Tubes for diluting samples. 4. Deionized or distilled water. 5. Absorbent paper for blotting the microtiter plate. 6. Container for Wash Solution Sample Preparation and Storage: Serum: Allow samples to clot for two hours at room temperature or overnight at 4C before centrifugation for 20 minutes at approximately 1000xg. Assay freshly prepared serum immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Plasma: Collect plasma using EDTA or heparin as anticoagulant. Centrifuge samples for 15 minutes at 1000xg at 2-8C within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquots at -20C or -80C for later use. Avoid repeated freeze/thaw cycles. Note: Serum/plasma samples require abou 100- fold dilution. A suggested 100-fold dilution is: 10ul sample + 990ul PBS. Samples should be diluted using 0.01M PBS (pH 7.0-7.2). Cell Lysates: Cells must be lysed before assaying according to the following directions. 1. Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly). 2. Wash cells three times in cold PBS. 3. Resuspend cells in PBS (1*) and subject the cells to ultrasonication 4X (or freeze cells at -20C. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle 3X.) 4. Centrifuge at 1500*g for 10 minutes at 2-8C to remove cellular debris. Other Biological Fluids: Centrifuge samples for 20 minutes at 1000*g. Remove particulates and assay immediately or store samples in aliquots at -20C or -80C. Avoid repeated freeze/thaw cycles. Note: 1. Samples to be used within 5 days may be stored at 4C, otherwise samples must be stored at -20C (1 month) or -80C (2 months) to a