Liver tissue was isolated and snap-frozen in liquid nitrogen and stored at -80°C. The biochemical determination of liver cholesterol and triglyceride levels were carried out as described previously (7 (link)). Briefly, 50 mg of frozen liver tissue was homogenized for 30 seconds at 5000 rpm in a closed tube with 1.0-mm glass beads and 1.0 ml SET buffer (Sucrose 250 mmol/l, EDTA 2 mmol/l and Tris 10 mmol/l). Complete cell destruction was done by 2 freeze-thaw cycles and 3 times passing through a 27-gauge syringe needle and a final freeze-thaw cycle. Protein content was measured using the BCA method Pierce, Rockford, IL). Liver cholesterol and triglyceride levels were quantified using cholesterol liquicolor kit [CHOD-PAP, Roche, Basel, Switzerland; triglycerides: GPOtrinder, TRO100, Sigma Aldrich, St. Louis, MO, USA)] following the manufacturer’s protocol on a Benchmark 550 micro-plate reader (Bio-Rad, Hercules, CA, USA).
Benchmark 550 micro plate reader
Manufactured by Bio-Rad
Sourced in United States
The Benchmark 550 micro-plate reader is a compact and versatile instrument designed for absorbance-based assays. It features a high-performance monochromator and a sensitive photodetector, enabling accurate and reliable measurements across a wide range of wavelengths. The Benchmark 550 is capable of performing quantitative and qualitative analyses in various applications, including enzyme-linked immunosorbent assays (ELISA), cell-based assays, and other plate-based experiments.
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Lab products found in correlation
3 protocols using benchmark 550 micro plate reader
1
Quantification of Liver Lipid Levels
2
Plasma and Liver Lipid Analyses
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Total plasma cholesterol and triglyceride levels were measured (1489232, Chol CHOD-PAP, Roche, Almere, the Netherlands; 337-B, TG GPO-trinder, Sigma Aldrich, Zwijndrecht, the Netherlands). Measurements were done according to manufacturer's protocols on a Benchmark 550 Micro-plate Reader (170-6750XTU, Bio-Rad, Veenendaal, the Netherlands). For organ lipid analyses, ~50 mg of frozen liver was homogenized for 30 s at 5,000 rpm in a closed tube with 5.0 mm glass beads and 1.0 ml SET buffer (Sucrose 250 mM, EDTA 2 mM, and Tris 10 mM) (35 (link)). Complete cell destruction was done by two freeze-thaw cycles and 3 times passing through a 27- gauge syringe needle and a final freeze-thaw cycle. Protein content was measured with the BCA method (23225, Pierce, Rockford, IL, United States). TG were measured as described above. All analyses were performed according to manufacturers' instructions.
3
Plasma Biomarkers and Body Composition
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After overnight fasting, venous blood was collected into EDTA tubes that were put on ice after blood collection at Maastricht University Medical Center (MUMC+). After centrifuging (1000 x g; 10 min; 4°C), plasma was snapfrozen in liquid nitrogen and then stored at -80°C until analyses. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), bilirubin, Alkaline Phosphatase, total cholesterol, low density lipoprotein cholesterol (LDL-cholesterol), high density lipoprotein cholesterol (HDL-cholesterol), triglyceride, fasting plasma glucose and hemoglobin A1c (HbA1C) were determined using routine analyses at the clinical chemistry department of the Maastricht UMC+ hospital. Plasma CTSD levels were determined using the CTSD enzyme-linked immunosorbent assay according to the manufacturer's protocol (Uscn Life Science, Wuhan, China). The absorbance for CTSD levels was measured on a Benchmark 550 microplate reader (Bio-Rad, Hercules, CA). Skeletal muscle mass index (SMI) (appendicular skeletal mass/height 2 ) were calculated with dual-energy X-ray absorptiometry (DXA).
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