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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.


Images

Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (AB105134), expandable thumbnail
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (AB105134), expandable thumbnail
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (AB105134), expandable thumbnail
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (AB105134), expandable thumbnail
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (AB105134), expandable thumbnail

Publications

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)
Reactive species
Mammals
Assay time
1h 20m
Sensitivity
> 10 mU/well

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Target data

Function

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).

Alternative names

What's included?

2000 Test
Components
ALT Positive Control
20 x 1 Vial
ALT Substrate Mix
20 x 1 Vial
Assay Buffer 13
20 x 25 mL
Developer Mix A
20 x 1 Vial
OxiRed™ Probe
20 x 0.2 mL
Pyruvate Standard
20 x 100 µL

Recommended products

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.

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)
Reactive species
Mammals
Assay time
1h 20m
Assay Platform
Microplate reader
Sensitivity
> 10 mU/well

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Associated diseases and disorders

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.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail
    Waller-Evans H et al., PLoS One, 8(12). Fig 1d. doi: 10.1371/journal.pone.0082825 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    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.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail
    Qiu Guihua, et al., Frontiers in Immunology Vol 8. 2017 pg.575, Fig 8, doi:10.3389/fimmu.2017.00575

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    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.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    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.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Pyruvate measured colorimetrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    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.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Pyruvate measured in biological fluids after 20 min and 40 min incubation time showing quantity (nmol) per ml of tested sample.

  • Enzyme activity assay - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Enzyme activity assay - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    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.

  • Biochemical assay - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134), expandable thumbnail

    Biochemical assay - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

    Diagram showing the principles of the ALT assay method.

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