Description
Introduction: alpha-smooth muscle actin (alpha-SMA) is expressed primarily in normal adult smooth muscle and transiently during development of cardiac and skeletal muscle. Expression, however, is not exclusive to smooth muscle cells, and is a prominent feature of other cells such as myofibroblasts. It is also the most commonly used marker gene for myofibroblast differentiation, which appear de novo at the early fibroproliferative stages of wound healing, desmoplastic responses in cancers, tissue repair/remodeling, and fibrosis in multiple organs.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to alpha-SMA. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for alpha-SMA and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain alpha-SMA, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The 3 enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of alpha-SMA in the samples is then determined by comparing the O.D. of the samples to the standard curve