Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 is a quantitative, 80-min ALT activity assay. In the assay, ALT acts to produce pyruvate, which is detected in a further reaction. Readout on any colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.
- Complete kit including standard curve for quantitation
- Over 150 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Catalyzes the reversible transamination between alanine and 2-oxoglutarate to form pyruvate and glutamate. Participates in cellular nitrogen metabolism and also in liver gluconeogenesis starting with precursors transported from skeletal muscles (By similarity).
Gpt1, Gpt, Alanine aminotransferase 1, ALT1, Glutamate pyruvate transaminase 1, Glutamic--alanine transaminase 1, Glutamic--pyruvic transaminase 1, GPT 1
Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 is a quantitative, 80-min ALT activity assay. In the assay, ALT acts to produce pyruvate, which is detected in a further reaction. Readout on any colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader.
- Complete kit including standard curve for quantitation
- Over 150 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 is a rapid and simple assay used to quantify alanine transaminase (ALT) activity in mammalian samples.
The kit has a detection limit of 10 mU per well.
Alanine transaminase is also called alanine aminotransferase or serum glutamic pyruvic transaminase (ALT, ALAT, SGPT).
How the assay works
In the ALT assay protocol, ALT transfers an amino group from alanine to α-ketoglutarate; producing pyruvate and glutamate. The pyruvate is detected in a reaction that converts a nearly colorless probe to a form that is colored (ODmax = 570 nm) and fluorescent (Ex/Em = 535/587 nm).
ALT assay protocol summary
ALT assay methods
There are 2 established ALT assay methods. In both methods, ALT transfers an amino group from alanine to alpha-ketoglutarate; producing pyruvate and glutamate.
Then in the preferred pyruvate oxidase method used in ALT Assay Kit ab105134, pyruvate oxidase acts on the pyruvate, producing hydrogen peroxide. This is then followed by the oxidation of a substrate by a peroxidase using the hydrogen peroxide to produce a colorimetric or fluorometric readout.
In the alternate LDH method, LDH acts on the pyruvate to produce lactate, in the process transforming NADH to NAD. Detection is by the decrease in absorbance by NADH at 340 nm.
A further method exists, where phenylhydrazine reacts with the pyruvate to form phenylhydrazone. Phenylhydrazone is a reddish brown solution under alkaline conditions, detected by absorbance at 510 nm.
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K752 Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric/Fluorometric Assay Kit. K752-100 is the same size as the 100 test size of ab105134.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
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Alanine transaminase also known as ALT or alanine aminotransferase is an important enzyme in the body. It has a molecular mass of approximately 54 kDa and catalyzes the conversion between alanine and pyruvate by transferring an amino group. ALT is mainly expressed in the liver but also present in other tissues like the heart and muscles. It is often measured as 'ALT activity' in clinical settings to ascertain liver health. In laboratories an 'ALT assay' or 'ALT kit' can determine alanine transaminase activity.
Alanine transaminase plays a critical role in amino acid metabolism and energy production. It facilitates the transamination process which is significant for the balance of amino acids and carbohydrates. ALT as a standalone enzyme does not form part of a protein complex but works closely with alanine aminotransferase reactions. It helps maintain the cellular nitrogen balance linking nitrogen metabolism with energy pathways.
The role of alanine transaminase is to interconnect carbohydrate and amino acid metabolism. It participates in the gluconeogenesis pathway and the citric acid cycle important for energy production. Through its activity in these pathways it interacts with proteins like aspartate aminotransferase (AST) affecting the balance of amino and keto acids in cells.
Alanine transaminase levels indicate liver conditions such as hepatitis and liver cirrhosis. Elevated ALT activity suggests liver damage or inflammation. It also connects to conditions involving muscular injury. In these scenarios increased presence and activity of AST often accompany ALT providing a broader picture of the extent of tissue damage. Understanding the involvement of ALT in these pathways helps diagnose and monitor the progression of relevant health issues.
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Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.
Liver samples from high fat diet (HFD) and standard carbohydrate diet (CHD) BALB/c and C57BL6/J mice were homogenised in an ALT assay buffer for the determination of ALT activity using ab105134. A separate batch of liver extracts was prepared and incubated in a buffer containing NP40 (5%) and supernatants containing the triglycerides were separated. Triglycerides concentration was determined on the supernatant fraction using Triglyceride Assay Kit - Quantification ab65336. ALT activity and triglycerides concentration were determined by measuring OD at 570nm.
Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
Serum aspartate transaminase (AST) levels were measured using Aspartate Aminotransferase Activity Assay Kit ab105135 and alanine transaminase (ALT) levels were measured using ab105134. Both levels were measured at various time periods post Con A injection. Mean values ± SD are shown (n = 4). ∗P < 0.05 and ∗∗P < 0.01.
Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
Alanine transaminase measured in mouse tissue lysates showing quantity (mU) per mg of tested sample.
Protein concentration for samples varied from 4 mg/mL to 13 mg/mL. Samples were diluted 9-27 fold and measured colorimetrically.
Pyruvate measured colorimetrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.
Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).
Functional Studies - Alanine Transaminase Activity Assay Kit (105134).
Pyruvate measured fluorometrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.
Pyruvate measured in biological fluids after 20 min and 40 min incubation time showing quantity (nmol) per ml of tested sample.
Alanine Aminotransferase assay kit ab105134 and Aspartate Aminotransferase assay kit Aspartate Aminotransferase Activity Assay Kit ab105135 used with mouse serum.
Hwang et al used Alanine Aminotransferase assay kit ab105134 and Aspartate Aminotransferase assay kit Aspartate Aminotransferase Activity Assay Kit ab105135 to investigate the impact of endothelial-specific-TAZ-knockout of mice fed with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) to induce fibrosis.
Partial hepatectomy (PHx) was conducted in WT and eKO mice, following which the livers were subjected to Sirius Red staining to visualize fibrosis, at 8 d after PHx. Provided data are representative images from multiple mice. The stained area was measured and analyzed using ImageJ (n = 4). Serum of each mouse was analyzed to determine the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity (n = 8 for ALT and n = 6 for AST).
Mouse serum was prepared by collecting blood by means of tail bleeding, followed by centrifugation after blood clotting, and incubated at room temperature for 30 min. The supernatant was collected and directly assessed using an Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric/Fluorometric Assay Kit (K752, BioVision, now manufactured by Abcam as ab105134). The subsequent experimental steps were performed according to the manufacturer's protocol.
Aspartate aminotransferase (AST) activity assay: Mouse serum was prepared by tail bleeding. The isolated serum was assayed directly using an aspartate aminotransferase (AST) activity assay kit (Aspartate Aminotransferase Activity Assay Kit ab105135, Abcam). Subsequent experimental steps were performed according to the manufacturer's protocol.
Diagram showing the principles of the ALT assay method.
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