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Total cholesterol

Manufactured by Thermo Fisher Scientific
Sourced in Japan, Canada

The Total Cholesterol assay is a laboratory test used to measure the total amount of cholesterol present in a blood sample. It provides a comprehensive assessment of an individual's cholesterol levels, which is an important indicator of cardiovascular health.

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9 protocols using total cholesterol

1

Plasma Metabolite Quantification Protocol

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Plasma insulin levels were determined using an ELISA kit (Crystal Chem, Downers Grove, IL). Plasma levels of total cholesterol (Thermo Electron, Louisville, CO) and free and total glycerol (Sigma) were determined using commercial colorimetric assays. Plasma true triglyceride concentrations were estimated by subtraction of free glycerol from total glycerol.
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2

Plasma Lipid and Cytokine Analysis

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Blood was collected when animals were euthanized at 17-18 weeks of age and plasma obtained by centrifugation (1,200g) at 4°C. Plasma levels of total cholesterol (Thermo Electron, Louisville, CO) and free and total glycerol (Sigma, St. Louis, MO) were determined using colorimetric assays. Plasma true triglyceride concentrations were estimated by subtraction of free glycerol from total glycerol. Plasma IL6 and Insulin levels were determined using an ELISA kit according to manufacturer’s instructions (IL6, R&D System, Minneapolis, MN; Insulin, Crystal Chem, Downers Grove, Illinois).
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3

Multiparametric Metabolic Profiling

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Plasma glucose, alanine amino transferase (ALT), uric acid, total cholesterol, HDL cholesterol (Thermo Electron, Pittsburgh, PA), and NEFA (WAKO Pure Chemical Industries Ltd., Osaka, Japan) were determined using a Konelab 20X (Thermoscientific, Vantaa, Finland). Plasma leptin and insulin concentrations were measured using commercially available immunoassay kits (Crystal Chem Inc, Downers Grove, IL, USA and ALPCO Diagnostics, Salem, NH, USA).
All assays were conducted in accordance with the manufacturer's instructions and intra-and inter-assay coefficients of variation were always <10%.
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4

Enzymatic Assay for Lipid Profiling

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Enzymatic assay kits were used to measure total cholesterol (Thermo Fisher, Ottawa, ON, Canada), unesterified cholesterol, and HDL-cholesterol (FujiFilm Medical Systems, formerly Wako Diagnostics, Richmond, VA) following the manufacturer’s instructions. Cholesteryl ester and non-HDL cholesterol values were determined by subtraction of unesterified cholesterol or HDL-cholesterol (respectively) from total cholesterol.
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5

Plasma Biomarker Measurement Protocol

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Plasma levels of alanine transaminase (ALT) and aspartate transaminase (AST) were determined using kinetic absorbance assays (Teco Diagnostics) as described previously [16 (link)]. Analysis of plasma triglycerides (Thermo Fisher Scientific), total cholesterol (Thermo Fisher Scientific), and non-esterified fatty acids (FUJIFILM Wako) were determined using colorimetric based assay as previously reported [50 (link), 51 (link)].
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6

Comprehensive Metabolic Biomarker Assessment

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Immunoassays were used to measure total GIP (Millipore, ELISA), total GLP-1, and insulin (Meso Scale Discovery). Enzymatic assays were used to measure TG, beta-hydroxybutarate (Sigma-Aldrich), and total cholesterol (Thermo Fisher). HDL-cholesterol was determined in the supernatant after removal of LDL/vLDL-cholesterol by a precipitation reagent (Sigma-Aldrich); non-HDL-cholesterol was determined by subtracting HDL from total cholesterol. Free fatty acids were measured by GC-MS.
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7

Lipid Quantification in Liver Samples

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Liver samples (0.1 g) were homogenized in 2 ml Folch (chloroform/methanol, 2:1, v/v) with a polytron homogenizer. The organic phase was separated and then dried under nitrogen. Samples were reconstituted in isopropranol:Triton-X100 (9:1 v/v) and aliquots subjected to colorimetric enzymatic assays for total cholesterol (Thermo Scientific) or triglycerides (Pointe Scientific).
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8

Synthesis and Characterization of SR-135 Compounds

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SR-135 and its control compound SRB were synthesized as previously reported (31 (link)). Collagenase type XI, Hanks’ balanced salt solution, and monoclonal mouse anti-glucagon antibody were obtained from Sigma (St. Louis, MO). Total cholesterol, HDL-cholesterol, and triglyceride liquid reagents and their respective standards, fetal bovine serum, and penicillin-streptomycin were from Fisher Scientific (Pittsburgh, PA) and CMRL-1066 was from Invitrogen (Grand Island, NY). Insulin rabbit mAb and cleaved caspase 3 rabbit mAb were obtained from Cell Signaling (Danvers, MA). The secondary antibodies, DyLight 649-conjugated donkey anti-rabbit or anti-mouse IgG and Alexa 488-conjugated donkey anti-rabbit or anti-mouse antibody were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA) and 4′-6-diamidino-2-phenylindole (DAPI) were obtained from Invitrogen (Grand Island, NY). Immunocruz rabbit ABC staining system was from Santa Cruz Biotechnology (Dallas, Texas) and 3-nitrotyrosine mouse mAb was from Abcam (Cambridge, MA). Insulin mouse mAb and rat insulin RIA kit was from Millipore (St. Louis, MO). All other chemicals were from commercially available sources.
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9

Rodent Diet and Biomarker Analysis

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Laboratory Rodent Diet 5001 was purchased from EL Mel (St. Louis, MO) and Sucrose Dustless Precision Pellets were from Bio-Serve (Flemington, NJ). Insulin rabbit mAb was obtained from Cell Signaling (Danvers, MA). Stretozotocin and monoclonal mouse anti-glucagon antibody were obtained from Sigma (St. Louis, MO). The secondary antibodies, Alexa 647-conjugated donkey anti-rabbit antibodies and Alexa 488-conjugated donkey anti-mouse were obtained from Jackson Immuno Research Laboratories (West Grove, PA). Schiff reagent, periodic acid, total cholesterol, high density lipoprotein (HDL)-cholesterol, and triacylglyceride (TAG) liquid reagents and their respective standards were purchased from Fisher Scientific (Pittsburgh, PA) and alpha-amylase was from MilliporeSigma (Burlington, MA). Contour Next test strips were obtained from ADW Diabetes (Pompano Beach, FL). Rat hemoglobin A1c kit and control for the kit were obtained from Crystal Chem (Elk Grove Village, IL).
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