All chemicals were purchased from Sigma Chemical Co. (St. Louis, MO) unless stated otherwise. Mice were intraperitoneally (i.p.) injected with 300 mg/kg APAP (dissolved in warm saline) or saline vehicle after overnight fasting. The animals were sacrificed 6, 12, 24, 48 or 72 h after APAP treatment, blood was withdrawn from the vena cava into a heparinized syringe for measurement of alanine aminotransferase (ALT) activities (Pointe Scientific, Canton, MI) and flow cytometric analysis. The liver was removed and rinsed in saline; liver sections were fixed in 10% phosphate buffered formalin for histological analyses. The remaining liver was used for isolation of non-parenchymal cells (as described below). Additionally, as a positive control, some mice were treated (i.p.) with 100 μg/kg Salmonella abortus equi endotoxin (ET) for 90 min and then sacrificed (approximately at 10 am).
Alanine aminotransferase
Manufactured by Horiba
Sourced in United States
Alanine aminotransferase is an enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, forming pyruvate and glutamate. It is commonly used as a biomarker in the detection and monitoring of liver diseases.
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2 protocols using alanine aminotransferase
1
Acetaminophen-Induced Liver Injury in Mice
2
Liver Function Markers and Histology
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Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) assay kits were purchased from Pointe Scientific (Canton, MI, USA). A bile acid assay kit was purchased from Diazyme Laboratories (Poway, CA, USA). Glycine-conjugated β muricholic acid (G-β-MCA) was purchased from MedChemExpress Inc. (Monmouth Junction, NJ, USA). F4/80 antibody (Cat #. 70076) was purchased from Cell Signaling Technology (Danvers, MA, USA). CK19 antibody (ab52625) was purchased from Abcam (Waltham, MA, USA). ZO-1 antibody (PA5-28858) was purchased from ThermoFisher Scientific (Grand Island, NY, USA). Fluorescein isothiocyanate–dextran (FITC–dextran, MW:3000–5000) was purchased from Sigma Aldrich (St. Louis, MO, USA).
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