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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
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Add samples and standards to wells
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Add reaction mix and incubate for 10 min at 37°C
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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. 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. |
