Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric/Fluorometric
- Platform: Microplate reader
- Assay time: 1 hr 20 min
- Sample type: Cell culture media, Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 10 mU/well
Overview
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Product name
Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) -
Detection method
Colorimetric/Fluorometric -
Sample type
Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media -
Assay type
Enzyme activity (quantitative) -
Sensitivity
> 10 mU/well -
Assay time
1h 20m -
Product overview
Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 is a rapid and simple assay used to quantify alanine transaminase (ALT) activity in mammalian samples.
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).
The kit has a detection limit of 10 mU per well.
ALT assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 10 min at 37ºC
- analyze every 2-3 min for 60 min with microplate reader in kinetic mode at 37ºC -
Notes
Alanine transaminase is also called alanine aminotransferase or serum glutamic pyruvic transaminase (ALT, ALAT, SGPT).
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests ALT Positive Control (lyophilized) Blue 1 vial ALT Substrate (lyophilized) Orange 1 vial Pyruvate Standard (100 nmol/µl) Yellow 1 x 100µl ALT Assay Buffer WM 1 x 25ml ALT Enzyme Mix (lyophilized) Green 1 vial OxiRed™ (in DMSO Red 1 x 200µl -
Research areas
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Alternative names
- Alanine Aminotransferase
- ALT
Images
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Determination of ALT activity using ab105134 Alanine Transaminase Activity Assay Kit 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/
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 ab65336. ALT activity and triglycerides concentration were determined by measuring OD at 570nm.
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Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted. -
Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134) Qiu Guihua, et al., Frontiers in Immunology Vol 8. 2017 pg.575, Fig 8, doi:10.3389/fimmu.2017.00575Serum aspartate transaminase (AST) levels were measured using 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
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Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
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Functional Studies - Alanine Transaminase Activity Assay Kit (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.
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Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)Pyruvate measured colorimetrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.
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Functional Assay - ab105134Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).
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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.
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Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)Pyruvate measured in biological fluids after 20 min and 40 min incubation time showing quantity (nmol) per ml of tested sample.
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Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.
