Circulating fasting plasma glucose levels (Infinity Glucose Hexokinase, Thermo Fisher), total cholesterol (Infinity Cholesterol, Thermo Fisher), triglycerides (Infinity Triglycerides, Thermo Fisher), HDL-C (Infinity Cholesterol, Thermo Fisher), and non-esterified free fatty acids (Wako NEFA-HR(2 (link)), Wako Diagnostics) were quantified with in vitro enzymatic colorimetric method assays. FSH, insulin, TNFα, and IL6 concentrations were determined using a MILLIPLEX magnetic bead-based quantitative multiplex immunoassay with the MAGPIX instrumentation (Millipore, Billerica, MA).
Infinity cholesterol
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Infinity Cholesterol is a laboratory instrument used for the quantitative determination of cholesterol levels in biological samples. It provides accurate and reliable measurements to support clinical and research applications.
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Lab products found in correlation
Infinity triglyceride, by Thermo Fisher Scientific (6 mentions)
Enzyme immunoassay kits, by Mercodia (2 mentions)
Infinity glucose hexokinase liquid stable reagent, by Thermo Fisher Scientific (2 mentions)
Hdl cholesterol plus direct, by Thermo Fisher Scientific (2 mentions)
Mouse lbp elsa kit, by Hycult Biotech (2 mentions)
β hydroxybutyrate, by Thermo Fisher Scientific (2 mentions)
Non esterified fatty acid, by Fujifilm (2 mentions)
Infinity triglyceride reagent, by Thermo Fisher Scientific (2 mentions)
Infinity triglycerides measurement, by Thermo Fisher Scientific (2 mentions)
Hr series nefa hr 2, by Fujifilm (1 mentions)
Trizoltm, by Thermo Fisher Scientific (1 mentions)
Autokit glucose, by Fujifilm (1 mentions)
Phospholipids c, by Fujifilm (1 mentions)
Qpcrtm core kit for sybr green 1, by Eurogentec (1 mentions)
14c oleic acid, by PerkinElmer (1 mentions)
14c triolein, by PerkinElmer (1 mentions)
3h cholesterol, by PerkinElmer (1 mentions)
Omniscript rt, by Qiagen (1 mentions)
Poloxamer 407, by Spectrum Chemical (1 mentions)
Mb100b, by R&D Systems (1 mentions)
17 protocols using infinity cholesterol
1
Metabolic and Inflammatory Biomarkers
2
Metabolic and Inflammatory Biomarkers
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Fasting glucose, total, and high-density lipoprotein (HDL) cholesterol levels were measured in plasma using Infinity™ Glucose Hexokinase Liquid Stable Reagent, Infinity™ Cholesterol, and HDL-CHOLESTEROL Plus Direct method, respectively (Thermo Fisher Scientific Inc., Waltham, MA, USA). Insulin levels were quantified by enzyme immunoassay kits (Mercodia AB, Uppsala, Sweden). Lipopolysaccharide binding protein (LBP) was measured by mouse LBP ELSA kit (Hycult Biotech Inc., Wayne, NJ, USA). Serum amyloid A (SAA) levels were determined by “PHASE” ™ Murine Serum Amyloid A Assay (Tridelta Development Ltd., Maynooth, Ireland). Cytokine levels were measured in plasma (dilution 1:1) using Meso Scale Discovery V-PLEX proinflammatory panel one kit (#K15048D-1) as described in section 2.8. In this case, the calculated detection limits were the following: IFN-γ (0.00988–909 pg/mL), IL-1β (0.374–1,765 pg/mL), IL-2 (0.194–2,595 pg/mL), IL-4 (0.0266–1,680 pg/mL), IL-5 (0.224–1,027 pg/mL), IL-6 (0.335–5,440 pg/mL), IL-10 (0.103–3,930 pg/mL), IL-12p70 (1.79–31,400 pg/mL), KC/GRO (0.0968–1,960 pg/mL), TNF-α (0.0256–619 pg/mL).
3
Comprehensive Lipid Metabolism Profiling
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Infinity Cholesterol (catalog #TR13421), Infinity Triglyceride (catalog #TR22421), and TRIzolTM (catalog #15596018) reagents were purchased from Thermo Fisher Scientific (Middletown, VA, USA). Autokit Glucose (catalog #997-03001), Phospholipids C (catalog #997-01801), and HR Series NEFA-HR(2) (catalog #999-34691, 995-34791, 991-34891, and 993-35191) kits were purchased from Fujifilm Wako Chemicals USA (Richmond, VA, USA). Omniscript RT (catalog #205113) kit was purchased from Qiagen (Germantown, MD, USA) and qPCRTM core kit for SYBR Green I (catalog #10-SN10-05) was from Eurogentec (San Diego, CA, USA). 3H-cholesterol (catalog #NET139001MC), 14C-oleic acid (catalog #NEC317250UC), and 14C-triolein (catalog #NEC674250UC) were from PerkinElmer (Shelton, CT, USA). Poloxamer 407 (catalog #P1166) was purchased from Spectrum Chemical (New Brunswick, NJ, USA). Primary and secondary antibodies were purchased from either Cell Signaling (Danvers, MA, USA) or Abcam (Cambridge, MA, USA). All other chemicals and solvents were obtained from Fisher Scientific through its local distributor in the Kingdom of Saudi Arabia.
4
Liver Lipid Biomarker Measurement Protocol
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Liver triglycerides were measured using an enzymatic assay (#T7532-120, Pointe Scientific, Canton MI). Plasma was stored at -80 °C until further processing. Plasma was diluted 1:20 in saline in order to measure triglycerides (#TR22421, Infinity Triglyceride Reagent, Thermo Scientific, Waltham, MA). Cholesterol (Infinity Cholesterol, #TR13421, Thermo Scientific, Waltham, MS), total bile acids (#BQ 092A-EALD, BQkits Diagnostics, San Diego, CA), β-hydroxybutyrate (#SBHR-100, Fisher Scientific, Waltham, MA), and non-esterified fatty acids (Wako Diagnostics, Richmond, VA) were measured using enzymatic assays. TGFβ measurements were made using a standard ELISA (#MB100B, R&D Systems, Minneapolis, MN). Estradiol and progesterone assays were performed by the Vanderbilt Hormone Assay and Analytical Services Core (Vanderbilt University, Nashville, TN).
5
Plasma Cholesterol Determination
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Total plasma cholesterol were determined using Infinity™ Cholesterol (#TR13421, Thermo Fisher Scientific).
6
Plasma Cholesterol Quantification Protocol
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Total plasma cholesterol was determined using an enzymatic, colorimetric assay kit (Infinity Cholesterol; Thermo Fisher), according to manufacturer’s instructions. Briefly, plasma was isolated by centrifugation and stored at −80 °C until assaying. For each assay plate, a standard curve ranging between 0 and 300 mg dL−1 was run. A volume of 10 μL of standard or sample was pipetted onto a 96-well plate and 200 μL of working reagent (Infinity Cholesterol) added. The plates were covered, incubated for 15 min, and read at 550 nM (with a reference wavelength of 650 nM). All samples were performed in triplicate, where any replicates differing by 5% or more were re-assayed.
7
Blood Lipid Measurement in Mice
8
Plasma Lipid Analysis and Aortic Histology
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At the end of the feeding periods, blood was collected via the retro‐orbital sinus with the animals under anesthesia (intraperitoneal injection of ketamine [150 mg/kg] and xylazine [20 mg/kg]). Plasma was prepared by centrifugation of the collected blood samples at 2000g for 15 minutes and assayed for cholesterol and triglycerides using Infinity Cholesterol and Triglyceride Stable Reagents (Thermo Fisher Scientific, Middletown, VA). The mice were euthanized by cervical dislocation and immediately perfused with 30 mL of cold phosphate‐buffered saline via the right ventricle to remove blood components, following which the aortae were carefully dissected. The cleared aortae and lungs were flash‐frozen in liquid nitrogen for further biochemical analysis. Hearts (including aortic roots and ascending aorta) were removed, cut transversely, and embedded in optimum cutting temperature compound (Thermo Fisher Scientific). Serial sections (10‐µm thick) of ascending aortae were obtained for immunohistochemical analysis.
9
Metabolic and Inflammatory Biomarkers
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Fasting glucose, total, and high-density lipoprotein (HDL) cholesterol levels were measured in plasma using Infinity™ Glucose Hexokinase Liquid Stable Reagent, Infinity™ Cholesterol, and HDL-CHOLESTEROL Plus Direct method, respectively (Thermo Fisher Scientific Inc., Waltham, MA, USA). Insulin levels were quantified by enzyme immunoassay kits (Mercodia AB, Uppsala, Sweden). Lipopolysaccharide binding protein (LBP) was measured by mouse LBP ELSA kit (Hycult Biotech Inc., Wayne, NJ, USA). Serum amyloid A (SAA) levels were determined by “PHASE” ™ Murine Serum Amyloid A Assay (Tridelta Development Ltd., Maynooth, Ireland). Cytokine levels were measured in plasma (dilution 1:1) using Meso Scale Discovery V-PLEX proinflammatory panel one kit (#K15048D-1) as described in section 2.8. In this case, the calculated detection limits were the following: IFN-γ (0.00988–909 pg/mL), IL-1β (0.374–1,765 pg/mL), IL-2 (0.194–2,595 pg/mL), IL-4 (0.0266–1,680 pg/mL), IL-5 (0.224–1,027 pg/mL), IL-6 (0.335–5,440 pg/mL), IL-10 (0.103–3,930 pg/mL), IL-12p70 (1.79–31,400 pg/mL), KC/GRO (0.0968–1,960 pg/mL), TNF-α (0.0256–619 pg/mL).
10
Quantification of Plasma, Liver, and Fecal Lipids
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Plasma was recovered from heparinized blood samples after refrigerated centrifugation for 10 minutes at 2000 × g. Frozen liver tissues and vacuum-dried fecal samples were homogenized with ceramic beads (Bertin Technologies) prior to lipid content quantification. Plasma, liver, and fecal lipids were quantified through colorimetric enzymatic assays using Infinity triglycerides (Thermo Fisher Scientific) and Infinity cholesterol (Thermo Fisher Scientific) liquid-stable reagents (29 (link)) with signal detection using a microplate reader (Molecular Devices). Plasma glucose was quantified using a commercial kit (Wako Autokit Glucose 997-03001). Plasma monocyte chemotactic protein 1 (MCP-1) concentrations were determined using an ELISA kit (eBioscience, Inc.).
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