|Alanine Transaminase (ALT) Kit
Alanine Transaminase (ALT) Enzymatic Assay Kit is a rapid, plate-based test to measure the level of the liver enzyme Alanine transaminase (ALT), also known as Serum Glutamate Pyruvate Transaminase (SGPT) or Alanine Aminotransferase, in blood samples. Elevation of this enzyme in sera is indicative of liver damage and has been associated with liver injury, and toxicity associated with administration of toxic doses of experimental drugs and delivery agents. Therefore this enzyme provides a powerful means to examine chemically-induced liver damage. The convenient format of this simple, kinetic-based colorimetric test allows high-throughput screening of therapeutic pharmacophores, delivery agents and dosing schemes.
The MaxSuppressor™ Alanine Transaminase (ALT) Enzymatic Assay Kit uses a coupled enzymatic reaction scheme in which alanine and α-ketoglutarate are first converted to glutamate and pyruvate afterwhich the pyruvate is converted by lactate dehydrogenase to make lactate and NAD+. The conversion of the NADH chromophore to NAD+ product, measured at 340 nm, is proportional to the level of ALT enzyme in the sample. Each kit contains sufficient materials to perform 100 enzymatic reactions.
Convenient serum-based detection of acute hepato-toxicity caused by acetaminophen in mice using the BIOO Alanine Transaminase (ALT)Enzymatic Assay kit. Mice were administered with 0, 150 mg/kg, or 500 mg/kg dose of acetaminophen (intraperitoneal injection). 2 or 4 hours after injection, blood was collected from the mice. 10 μL of sera derived from these samples was mixed with assay reagent in 96 well plates in duplicate. The serum ALT enzymatic activity (a commonly used marker for liver organ damage) was detected using a UV-visible plate reader from the decrease in absorbance at 340 nm wavelength over a 5 minute interval. The plot shows the average absorbance decrease for each of the samples. A lower dose of acetaminophen (150 mg/kg) causes a slight increase in ALT activity while much higher elevation of ALT activity is observed with the higher dose (500 mg/kg).