The quantification of ALT activity in plasma was performed as indicated in the ALT Colorimetric Activity Assay kit (Cayman Chemicals, Ann Arbor, MI, USA).
Alt colorimetric activity assay kit
Manufactured by Cayman Chemical
Sourced in United States
The ALT Colorimetric Activity Assay Kit is a laboratory tool used to measure the activity of the enzyme Alanine Aminotransferase (ALT) in biological samples. The kit provides reagents and protocols to perform a colorimetric assay, allowing for the quantitative determination of ALT levels.
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Lab products found in correlation
Lactate plus meter, by Nova Biomedical (2 mentions)
Safire2 microplate reader, by Tecan (2 mentions)
Pathscan intracellular signaling array kit, by Cell Signaling Technology (1 mentions)
15 f2t isoprostane elisa kit, by Cayman Chemical (1 mentions)
Bsa protein assay, by Thermo Fisher Scientific (1 mentions)
Nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)
X tremegene hp, by Roche (1 mentions)
Tbars assay kit, by Cayman Chemical (1 mentions)
Mouse glucose assay kit, by Crystal Chem (1 mentions)
Ultra sensitive mouse insulin elisa kit, by Crystal Chem (1 mentions)
Bibf1120, by Selleck Chemicals (1 mentions)
β hydroxycyclodextrin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Simplyblue, by Thermo Fisher Scientific (1 mentions)
Mouse albumin elisa kit, by Abcam (1 mentions)
Hydroxyproline assay kit, by Cell Biolabs (1 mentions)
Infinity cholesterol kit, by Thermo Fisher Scientific (1 mentions)
Infinity triglyceride, by Thermo Fisher Scientific (1 mentions)
Total bilirubin reagent set, by Horiba (1 mentions)
Onetouch ultra, by LifeScan (1 mentions)
11 protocols using alt colorimetric activity assay kit
1
Quantifying Plasma ALT Activity
2
Reagents and Plasmids for Molecular Analyses
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Common cell culture media, supplements, culture plasticware, and commercial sources of protease inhibitors and DEX have been reported previously4 (link). The transfection reagent, X-tremeGENE HP was obtained from Roche (Indianapolis, IN) and SYBR green master mix from Lifescience Tech. (Carlsbad, CA); INH and Extract-N-Amp for mouse tissue genotyping from Sigma (St. Louis, MO); 15-F2t-Isoprostane ELISA kit, TBARS assay kit, ALT- colorimetric activity assay kit, and triglyceride (TG) detection assay from Cayman (Ann Arbor, MI), Oxyblot detection materials and EMSA gel shift assay from Millipore (Billerica, MA), Pathscan Intracellular Signaling array kit from Cell Signaling Technology (Danvers, MA), BSA protein assay, Pico ECL reagents, Nuclear and Cytoplasmic extraction reagents from Thermo Fisher (Pittsburgh, PA). pcDNA3-CHIP, pcDNA3-CHIP-∆U-box, pcDNA3-CHIP-∆TPR were provided by the Patterson lab. pEBG-AMPKα1 was provided by Dr. Rob Onyenwoke (North Carolina Central University). pCW45-AMPKα2 was obtained from DNA Resource Core of Harvard Medical School (Boston, MA).
3
Liver Injury and Metabolic Markers Assessment
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To analyse the liver injury and metabolic abnormalities in the liver, the levels of plasma alanine aminotransferase (ALT), plasma glucose, plasma insulin, plasma and liver triglyceride (TG), and liver cholesterol (TC) were determined. The plasma ALT was measured using an ALT Colorimetric Activity Assay kit (Cayman Chemical Company, Ann Arbor, MI, USA), according to the instructions of the manufacturer provided. Plasma insulin was measured using an Ultra Sensitive Mouse Insulin ELISA Kit, 90080 (Crystal Chem Inc, Downers Grove, IL, USA), according to the instruction provided. Plasma glucose assay was performed using a Mouse Glucose Assay Kit, 81692 (Crystal Chem Inc, Downers Grove, IL, USA). TG assay was conducted using a TG infinity assay kit (Thermo Fisher Scientific Inc, Waltham, MA, USA). Additionally, a TC assay was carried out using the TC infinity assay kit (Thermo Fisher Scientific Inc, Waltham, MA, USA).
4
Plasma ALT Measurement via Cayman Assay
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The levels of alanine transaminase (ALT) in plasma were measured using ALT colorimetric activity assay kit from Cayman Chemical Company (#700260) as per manufacturer's instructions.
5
Effects of BIBF1120 on CCl4-Induced Liver Injury
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To study the effect of BIBF1120, male C57BL/6 mice were treated with a single intraperitoneal injection of olive-oil or CCl4 (1 ml/kg in olive-oil) at day 1. At day 2 and day 3, CCl4-treated mice received intra-peritoneal administration of 5 mg/kg tyrosine kinase inhibitor BIBF1120 (Selleckchem, Boston, NY) prepared in 1% DMSO (Sigma) and 5% β-hydroxycyclodextrin (Sigma) or vehicle treatment (1%DMSO/5%β-hydroxycyclodextrin/PBS) (n = 5 per group). At day 4, all mice were sacrificed and livers were harvested for the subsequent analysis. Alanine Transaminase (ALT) levels in the plasma was measured using ALT Colorimetric Activity Assay Kit (Cayman Chemical Company, Michigan, MI, USA) as per manufacturer’s instructions.
6
Plasma Metabolite Analysis in Mice
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Retro-orbital bleeding was performed prior to sacrificing mice. Blood was collected in heparinized capillary tubes and centrifuged 10min at 7000 rpm at 4C. Supernatant plasma was saved for further serum analysis. Plasma lactate level was measured using a specific apparatus, Lactate Plus meter (Nova biomedical, cat 62624). ALT level was determined using ALT Colorimetric Activity Assay Kit (Cayman, cat 700260). Absorbances were detected using a Tecan safire2 microplate reader.
7
Liver and Kidney Dysfunction Biomarkers
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Serum alanine transaminase activity was analyzed as an indicator of liver dysfunction using a serum alanine transaminase (ALT) colorimetric activity assay kit (Cayman Chemical, USA). Blood urea nitrogen (BUN) levels were analyzed as an indicator of renal dysfunction (DetectX Urea Nitrogen Colorimetric Detection Kit, Funakoshi, Tokyo, Japan). Urinary proteins were analyzed by SDS-PAGE, followed by protein staining (Simply Blue, Invitrogen, Thermo Fisher Scientific). IL-6 and tumor necrosis factor (TNF)-α levels were evaluated to detect the systemic cytokine storm using a mouse IL-6 detection kit (Chondrex #6702, Woodinville, WA 98072, USA) and mouse TNF-α detection kit (Chondrex #6701), respectively, following the manufacturer’s protocols.
8
Plasma ALT Colorimetric Assay
9
Plasma Lactate and ALT Assay
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Retro-orbital bleeding was performed prior to sacrificing mice. Blood was collected in heparinized capillary tubes and centrifuged 10 min at 7000 rpm at 4°C. Supernatant plasma was saved for further serum analysis. Plasma lactate level was measured using a specific apparatus, Lactate Plus meter (Nova Biomedical, cat 62624). ALT level was determined using ALT Colorimetric Activity Assay Kit (Cayman, cat 700260). Absorbances were detected using a Tecan safire2 microplate reader.
10
Liver Enzyme and Albumin Assays
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