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
Product nameAlanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric)
See all Alanine Transaminase kits
Sample typeCell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media
Assay typeEnzyme activity (quantitative)
Sensitivity> 10 mU/well
Assay time1h 20m
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
Alanine transaminase is also called alanine aminotransferase or serum glutamic pyruvic transaminase (ALT, ALAT, SGPT).
Storage instructionsStore 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
- Alanine Aminotransferase
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.
Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
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.
Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
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.
Pyruvate measured colorimetrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.
Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).
Pyruvate measured fluorometrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.
Pyruvate measured in biological fluids after 20 min and 40 min incubation time showing quantity (nmol) per ml of tested sample.
Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.
ab105134 has been referenced in 63 publications.
- Humpton TJ et al. Differential requirements for MDM2 E3 activity during embryogenesis and in adult mice. Genes Dev 35:117-132 (2021). PubMed: 33334825
- Dong J et al. Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH. Nat Commun 12:66 (2021). PubMed: 33397952
- Hwang I et al. Retinol from hepatic stellate cells via STRA6 induces lipogenesis on hepatocytes during fibrosis. Cell Biosci 11:3 (2021). PubMed: 33407858
- Tajan M et al. Serine synthesis pathway inhibition cooperates with dietary serine and glycine limitation for cancer therapy. Nat Commun 12:366 (2021). PubMed: 33446657
- de la Rosa Rodriguez MA et al. Hypoxia-inducible lipid droplet-associated induces DGAT1 and promotes lipid storage in hepatocytes. Mol Metab 47:101168 (2021). PubMed: 33465519