Myostatin (GDF8, Growth Differentiation Factor 8, MSTN) is a member of the transforming growth factor beta superfamily that is essential for proper regulation of skeletal muscle mass. Myostatin is a secreted protein that negatively regulates skeletal muscle mass determining both muscle fiber number and size. The myostatin pathway is conserved and regulates muscle mass in a number of animal species ranging from fish to humans. Inhibition of myostatin using a variety of therapeutic approaches can increase muscle mass in a number of animal models of human disease, including muscular dystrophy.
Intended use:The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of MSTN in mouse serum, plasma and other biological fluids.
Test Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to MSTN. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for MSTN. 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 MSTN, 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 MSTN in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Precaution:M9875-11J: Stop Solution 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 *M9875-11A, *M9875-11B, *M9875-11D and *M9875-11E. Store at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial 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 2 hours at 37°C
3. Aspirate and add 100ul prepared Detection Reagent A immediately. Incubate 1 hour at 37°C. 4. 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 at 450nm immediately.
