Aspartate Transaminase Assay Kit

Details for Product No. ABIN1000292
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Target Name (Antigen)
Synonyms Afu6g02490, Afu2g09650
Detection Range 2 U/L.
Application
Activity Assay (AcA)
Quantity 100 tests
Shipping to United States (Change)
Availability Will be delivered in 4 to 6 Business Days
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Catalog No. ABIN1000292
295.90 $
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Sample Type Serum, Plasma, Biological Samples
Characteristics Sensitive. Linear detection range: 2-100 U/L.
Simple and convenient. This simple, convenient assay can be carried out in a microplate or a cuvette and takes only 10 min.
Components Assay Buffer: 24 mL. Cofactor: 120 µL. Enzyme Mix: 120 µL. NADH: 500 µL.
Material not included Pipeting devices and accessories. Clear bottom 96-well plates (e.g. Corning Costar) and plate reader or spectrophotometer and cuvettes for measuring OD340nm.
Alternative Name Aspartate Transaminase
Background Quantitative determination of aspartate transaminase activity at 340nm.
Procedure: 10 min.

Aspartate Transaminase (AST), also known as serum glutamic oxaloacetic transaminase (GOT) or aspartate aminotransferase (ASAT/AAT), facilitates the conversion of aspartate and alpha-ketoglutarate to oxaloacetate and glutamate. There are two isoenzymes in humans: GOT1 is a cytosolic isoenzyme derived from red blood cells and heart, GOT2 is the mitochondrial isoenzyme found mainly in the liver. AST is elevated in liver and muscle diseases. It is part of diagnostic tests for liver function, myocardial infarction, acute pancreatitis, acute hemolytic anemia, severe burns, acute renal disease and trauma. Simple, direct and automation-ready procedures for measuring AST activity find wide applications in research and drug discovery. This AST activity assay is based on the quantification of oxaloacetate produced by AST. In this assay, oxaloacetate and NADH are converted to malate and NAD by the enzyme malate dehydrogenase. The decrease in NADH absorbance at 340 nm is proportionate to AST activity.
Research Area Enzymes
Application Notes Direct Assays: AST activity in serum, plasma and other biological samples.
Drug Discovery/Pharmacology: effects of drugs on AST activity.
Protocol Procedure using 96-well plate
1.Samples and controls. Transfer 20 µL sample to each well. For each assay plate, include two wells with 20 µL distilled water to be used for the NADH Standard and Blank. Keep plate at the desired temperature (e.g. 37°C).
2.Prepare Working Reagent for Sample and Standard wells, by mixing for each well, 200 µL Assay Buffer, 1 µL Cofactor, 1 µL Enzyme Mix and 4 µL NADH. Warm to desired temperature (e.g. 37°C). Prepare Blank Reagent for the Blank well, by mixing 200 µL Assay Buffer, 1 µL Cofactor, 1 µL Enzyme Mix and 4 µL H2O. Warm to desired temperature (e.g. 37°C).
3. Add 200 µL Working Reagent to the Standard and Sample wells, and 200 µL Blank Reagent to the Blank well. Immediately tap plate to mix, incubate at the desired temperature and read OD340nm at 5 min and at 10 min. Alternatively, record kinetics at 340 nm.

Procedure using cuvettes
1. For each assay, include one Standard and one Blank control. For each Sample and Standard, prepare Working Reagent by mixing 1000 µL Assay Buffer, 5 µL Cofactor, 5 µL Enzyme Mix and 20 µL NADH. Transfer 990 µL Working Reagent to each sample cuvette and standard cuvette. Warm to desired temperature (e.g. 37°C). To Blank control cuvette, add 960 µL Assay Buffer, 5 µL Cofactor, 5 µL Enzyme Mix and 20 µL H2O. Warm to desired temperature (e.g. 37°C).
2. Prewarm sample to the desired temperature. Add 100 µL Sample to the Sample Cuvette. Transfer 100 µL H2O to the Standard cuvette and to Blank Control cuvette, respectively.
3. Mix immediately. Read OD340nm at 5 min and 10 min. Alternatively, record kinetics at 340 nm.
Reagent Preparation Equilibrate all components to room temperature. Keep thawed enzymes on ice. Assays can be performed at 37°C or at room temperature. Prior to assay, bring the working reagents, microplate and spectrophotometer to the desired temperature. Assay is compatible with serum or plasma (heparin, EDTA). Samples should be clear and free of particles or precipitates. Hemolyzed samples should not be used.
Restrictions For Research Use only
Storage -20 °C
Validation Images
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