Description
Introduction: Alanine transaminase or ALT is a transaminase enzyme (EC 2.6.1.2). It is also called serum glutamic pyruvic transaminase (SGPT) or alanine aminotransferase (ALAT). ALT is found in serum and in various bodily tissues, but is most commonly associated with the liver. It catalyzes the two parts of the alanine cycle. It catalyzes the transfer of an amino group from alanine to a-ketoglutarate, the products of this reversible transamination reaction being pyruvate and glutamate. Significantly elevated levels of ALT often suggest the existence of other medical problems such as viral hepatitis, congestive heart failure, liver damage, biliary duct problems, infectious mononucleosis, or myopathy. For this reason, ALT is commonly used as a way of screening for liver problems. However, elevated levels of ALT do not automatically mean that medical problems exist. Fluctuation of ALT levels is normal over the course of the day, and ALT levels can also increase in response to strenuous physical exercise. When elevated ALT levels are found in the blood, the possible underlying causes can be further narrowed down by measuring other enzymes.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to ALT. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for ALT 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 ALT, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The 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 ALT in the samples is then determined by comparing the O.D. of the samples to the standard curve