Total bile acids (TBA) were determined in serum and liver tissue using a TBA kit (Diazyme, Poway, CA) following manufacturer's instructions. In brief, 100 mg frozen liver tissue were minced and homogenized in 1 ml ice-cold phosphate-buffered saline with protease and phosphatase inhibitors (Pierce, Rockford, IL). Samples were centrifuged at 100,000g at 4 °C and the supernatants subjected to TBA determination. Liver cholesterol content was assayed using O-phthalaldehyde according to Rudel and Morris (Rudel and Morris 1973 (link)). Caspase-3 activity was detected in homogenized liver tissue using the Caspase-3 activity fluorometric assay kit (Calbiochem, Merck KGaA, Germany) and tyriglycerides were determined by the colorimetric triglyceride quantification kit (Abcam, Cambridge, MA) following manufacturer's instructions. Serum levels of cholesterol, aspartate aminotransferase (AST), alanine transferase (ALT), bilirubin (Bil), and alkaline phosphatase (ALP) were determined by automated method using Reflovet Plus (Roche Inc).
Reflovet plus
Manufactured by Roche
Sourced in Switzerland
The Reflovet Plus is a laboratory equipment product developed by Roche. It is a comprehensive solution for analyzing a range of clinical parameters in various sample types. The device provides accurate and reliable results to support clinical decision-making.
Automatically generated - may contain errors
Lab products found in correlation
Triglyceride colorimetric assay kit, by Cayman Chemical (3 mentions)
Reflotron got, by Roche (2 mentions)
Reflotron gpt, by Roche (2 mentions)
Microtainer k2e tubes, by BD (1 mentions)
Sevofluorane, by Abbott (1 mentions)
Taurine assay kit, by Cell Biolabs (1 mentions)
Cobas c311, by Roche (1 mentions)
17 protocols using reflovet plus
1
Quantifying Hepatic Bile Acids and Lipids
2
Plasma Lipid and Enzyme Analysis
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Plasma was centrifuged at 1000× g at 4 °C for 10 min and stored at −80 °C. The levels of TG, LDL, HDL, and total cholesterol were measured using commercial kits (Roche Molecular Biochemical, Pleasanton, CA, USA). Moreover, the plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assayed by the Reflovet Plus equipment (Roche, Basel, Switzerland) using commercial kits: Reflotron GOT, Reflotron GPT (Roche, Basel, Switzerland). TG levels in the liver were assayed using the Triglyceride Colorimetric Assay Kit (Cayman Chemical, Ann Arbor, MI, USA), according to the manufacturer’s instructions.
3
Effect of Anti-LPS Colostrum on Liver Enzymes
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Example 4
The inventors evaluated whether the liver enzyme levels, which indicate liver injury, of animals orally administered with [Figure (not displayed)]nti-LPS enriched colostrum-derived immunoglobulin preparation are improved due to the treatment. Levels of aspartyl transaminase (AST) and alanine aminotransferase (ALT) activities were determined by a clinical chemistry analyzer, Reflovet Plus (Roche Diagnostics, GmbH, Mannheim, Germany).
4
Serum ALT Analysis in Animal Model
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Blood samples were obtained from each animal. Serum levels of alanine aminotransferase (ALT) were determined by the automated method using Reflovet Plus (Roche).
5
Mouse Blood Serum ALT Assay
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Mice were sacrificed by CO2-euthanasia and blood was obtained by puncture of the heart with a 25 gauge needle on a 1 mL syringe. Blood samples were incubated first for 20 min at RT followed by incubation for 20 min at 4°C. Afterwards, samples were spun down (14000 rpm) at 4°C for 5 min and serum-containing supernatant was collected and stored at -20°C until further analysis. For analysis of the serum alanine transaminase (ALT) level, 32 μL serum was pipetted on a Refloton® test stripe and measured on the spectrophotometer Reflovet Plus (Roche).
6
Liver Injury Biomarkers Post-Transplant
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24h post-transplant aspartate aminotransferase (AST), alanine aminotransferase (ALT), levels were measured on serum from centrifuged blood (11,000 rpm for 7 minutes at 4 °C) using a multiple strip machine (RefloVet Plus, Roche, Basel, Switzerland). HMGB1 ELISA assay was performed according to manufacturers’ instructions (IBL International GMBH).
7
Blood Sampling and Biochemical Analysis
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For measurement of clinical chemistry parameters, 100–150 µl blood from the vena saphena lateralis was collected in heparin coated tubes (BD Microtainer K2E tubes, Becton, Dickinson and Company, Franklin Lakes, NJ). Alkaline phosphatase (ALP), Aspartate transaminase (GOT), Alanine transaminase (GPT) and Urea were measured in a Reflovet Plus (Roche Diagnostics, GmbH, D-68298, Mannheim, Germany). In some mice from experiment 4, blood samples were taken by heart puncture under gas anesthesia (Sevofluorane, Abbot, Abbot Park, IL).
8
Fasted Serum Lipid Profiling
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Triglyceride (TG) and total cholesterol serum levels were measured weekly in a fasted state using commercially available sticks with a Reflovet Plus clinical chemistry analyzer (Roche Diagnostics, GmbH, Mannheim, Germany).
9
Weekly ALT Serum Monitoring
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Serum alanine aminotransferase (ALT) was measured weekly using commercially available sticks (Reflovet Plus, Roche Diagnostic).
10
Evaluating Liver Enzyme Levels in Anti-LPS Colostrum Treatment
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Example 11
Having shown that oral administration of anti-LPS enriched colostrum-derived immunoglobulin preparation improves various metabolic syndrome markers, such as decreasing glucose tolerance and decreasing serum insulin, the inventors next evaluated whether the liver enzyme levels, which indicate liver injury, of animals fed with the preparation have also improved due to the treatment. Levels of AST and ALT activities were determined by a clinical chemistry analyzer, Reflovet Plus (Roche Diagnostics, GmbH, Mannheim, Germany).
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