Macrophage Migration Inhibitory Factor (MIF) BioAssay(TM) ELISA Kit (Mouse)

Produktübersicht

• Artikelname: Macrophage Migration Inhibitory Factor (MIF) BioAssay(TM) ELISA Kit (Mouse)
• Artikelnummer: USB-026649
• Hersteller Artikelnummer: 026649
• Alternativnummer: USB-026649-96
• Hersteller: US Biological
• Kategorie: Kits/Assays

Produktbeschreibung

The Mouse Macrophage Migration Inhibitory Factor (MIF) ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of MIF in mouse serum, plasma and other biological fluids. Detection Range: 31.2-2,000pg/ml Sensitivity: <14.6pg/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 MIF. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to MIF. 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 MIF, 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 MIF in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *026649A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *026649B: Standard, 2x1vial 026649C: Standard Diluent, 1x20ml *026649D: Detection Reagent A, 1x120ul *026649E: Detection Reagent B, 1x120ul 026649F: Assay Diluent A, 1x12ml 026649G: Assay Diluent B, 1x12ml 026649H: TMB Substrate, 1x9ml 026649K: Stop Solution, 1x6ml 026649L: Wash Buffer, 30x, 1x20ml Precaution: The Stop Solution (026649K) 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 *026649A, *026649B, *026649D and *026649E 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: Use a serum separator and 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 about 50-fold dilution (example: 10ul sample + 490ul PBS). Samples should be diluted using 0.01M PBS, pH 7.0 - 7.2. 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 avoid loss of bioactivity and contamination. 2. Sample hemolysis will influence the results so hemolyzed specimens should not be used. 3. When performing the assay, bring samples to RT. Assay Procedure Summary: 1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate 2 hours at 37C. 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C