Alanine Transaminase Activity Assay试剂盒(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 Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine (UTI)
- Sensitivity: 10 mU/well
概述
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产品名称
Alanine Transaminase Activity Assay试剂盒(Colorimetric/Fluorometric)
参阅全部 Alanine Transaminase 试剂盒 -
检测方法
Colorimetric/Fluorometric -
样品类型
Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media, Urine (UTI) -
检测类型
Enzyme activity (quantitative) -
灵敏度
> 10 mU/well -
检测时间
1h 20m -
种属反应性
与反应: Mammals -
产品概述
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. -
说明
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.The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
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平台
Microplate reader
性能
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存放说明
Store at -20°C. Please refer to protocols. -
组件 100 tests 2000 tests ALT Positive Control 1 vial 20 vials ALT Substrate Mix 1 vial 20 vials Assay Buffer XIII 1 x 25ml 20 x 25ml Development Enzyme Mix I 1 vial 20 vials OxiRed Probe 1 x 200µl 20 x 200µl Pyruvate Standard 1 x 100µl 20 x 100µl -
研究领域
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别名
- Alanine Aminotransferase
- ALT
图片
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Determination of ALT activity using ab105134 Alanine Transaminase Activity Assay KitWaller-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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Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
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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.2017.00575
Serum 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 < 0.05 and ∗∗P < 0.01.
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Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
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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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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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Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).
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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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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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Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.
数据表及文件
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SDS download
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Datasheet download
文献 (149)
ab105134 被引用在 149 文献中.
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- Bruno MEC et al. PAI-1 as a critical factor in the resolution of sepsis and acute kidney injury in old age. Front Cell Dev Biol 11:1330433 (2023). PubMed: 38304613
- Sarver DC et al. Dysregulated systemic metabolism in a Down syndrome mouse model. Mol Metab 68:101666 (2023). PubMed: 36587842
- Reis R et al. Alpha-Lipoic Acid Modulates the Oxidative and Inflammatory Responses Induced by Traditional and Novel Tobacco Products in Human Liver Epithelial Cells. Chem Biodivers 20:e202200928 (2023). PubMed: 36650104
- Liao JT et al. D-Limonene Promotes Anti-Obesity in 3T3-L1 Adipocytes and High-Calorie Diet-Induced Obese Rats by Activating the AMPK Signaling Pathway. Nutrients 15:N/A (2023). PubMed: 36678138