Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine (UTI), Plasma, Tissue Extracts, Cell culture media, Cell Lysate, Serum, Other biological fluids,
Assay type:Enzyme activity (quantitative),
Sensitivity:> 10 mU/well,
Assay time:1h 20m,
Assay Platform:Microplate reader
Product details:
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
- Add samples and standards to wells
- Add reaction mix and incubate for 10 min at 37°C
- Analyze every 2-3 min for 60 min with microplate reader in kinetic mode at 37°C
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.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
Biological function summary
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.
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.
