Triglyceride reagent

Manufactured by Horiba

The Triglyceride Reagent is a laboratory product designed to measure the concentration of triglycerides in biological samples. It provides a reliable and accurate method for the quantitative determination of triglycerides.

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Lab products found in correlation

Triglyceride standard, by Horiba (4 mentions) Clear flat bottom 96 well plate, by Corning (3 mentions) Softmax pro version 5, by Molecular Devices (1 mentions) Vmax microplate reader, by Molecular Devices (1 mentions) D12492, by LabDiet (1 mentions) 4 hne, by Cell Biolabs (1 mentions) Phospholipids c kit, by Fujifilm (1 mentions) Precimat glycerol, by Roche (1 mentions) Tissuelyser, by Qiagen (1 mentions)

12 protocols using triglyceride reagent

Quantification of Hepatic Lipids and Enzymes

Cited 2 times

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Hepatic triglycerides (TGs) were quantified using Triglyceride Reagent and Triglyceride Standards (Pointe Scientific). Serum alanine aminotransferase (ALT) and aspartate aminotransferase levels (AST) were quantified using ALT Reagent, AST Reagent and Catatrol I and II (Catachem). For histology, liver tissue was fixed in 10% buffered formalin, and stained with H&E or Masson’s trichrome and evaluated by a board-certified pathologist (3 (link), 24 (link)).

Oates J.R., Sawada K., Giles D.A., Alarcon P.C., Damen M.S., Szabo S., Stankiewicz T.E., Moreno-Fernandez M.E, & Divanovic S. (2023). Thermoneutral housing shapes hepatic inflammation and damage in mouse models of non-alcoholic fatty liver disease. Frontiers in Immunology, 14, 1095132.

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Quantifying Hepatic Triglycerides and Liver Function

Cited 4 times

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Hepatic triglycerides were quantified using Triglyceride Reagent and Triglyceride Standards (Pointe Scientific) as previously described (Giles et al., 2016 (link); Giles et al., 2017 ; Harley et al., 2014 (link); Moreno-Fernandez et al., 2018 ; Mukherjee et al., 2018 (link)). Serum alanine transaminase (ALT) levels were quantified using ALT Reagent and Catatrol I and II (Catachem). For histology, liver tissue was fixed in 10% buffered formalin, and stained with H&E. NAFLD activity score (NAS) was determined from H&E staining by a certified pathologist according to standard practice (Brunt et al., 2011 (link); Giles et al., 2016 (link); Giles et al., 2017 ; Harley et al., 2014 (link); Moreno- Fernandez et al., 2018 ).

Moreno-Fernandez M.E., Giles D.A., Oates J.R., Chan C.C., Damen M.S., Doll J.R., Stankiewicz T.E., Chen X., Chetal K., Karns R., Weirauch M., Romick-Rosendale L., Xanthakos S.A., Sheridan R., Szabo S., Shah A.S., Helmrath M.A., Inge T.H., Deshmukh H., Salomonis N, & Divanovic S. (2021). PKM2-dependent metabolic skewing of hepatic Th17 cells regulates pathogenesis of non-alcoholic fatty liver disease. Cell metabolism, 33(6), 1187-1204.e9.

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Hepatic Triglyceride and ALT Quantification

Cited 4 times

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Hepatic triglycerides were quantified using Triglyceride Reagent and Triglyceride Standards (Pointe Scientific) as previously described^{13 (link),22 (link)–24 (link),26 (link)}. Serum alanine transaminase (ALT) levels were quantified using ALT Reagent and Catatrol I and II (Catachem). For histology, liver tissue was fixed in 10% buffered formalin, and stained with H&E^{13 (link),22 (link),23 ,26 (link),27 (link)}.

Moreno-Fernandez M.E., Sharma V., Stankiewicz T.E., Oates J.R., Doll J.R., Damen M.S., Almanan M.A., Chougnet C.A., Hildeman D.A, & Divanovic S. (2021). Aging mitigates the severity of obesity-associated metabolic sequelae in a gender independent manner. Nutrition & Diabetes, 11, 15.

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Hepatic Triglyceride Quantification Protocol

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Hepatic TG levels were quantified as described.14, 17, 18 Briefly, standards and samples were added to a 96‐well, clear, flat‐bottom plate (Costar) containing 200 μL of triglyceride reagent (Pointe Scientific). Hepatic TGs were quantified at 500‐520 nm using the vmax Microplate Reader (Molecular Devices) and SoftMax Pro version 5 software.

Mukherjee R., Moreno‐Fernandez M.E., Giles D.A., Cappelletti M., Stankiewicz T.E., Chan C.C, & Divanovic S. (2018). Nicotinamide adenine dinucleotide phosphate (reduced) oxidase 2 modulates inflammatory vigor during nonalcoholic fatty liver disease progression in mice. Hepatology Communications, 2(5), 546-560.

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High-Fat Diet Induced Metabolic Dysregulation

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Diets: Mice were fed either a high-fat diet (HFD; Research Diets #D12492) or a chow diet (Chow; LAB Diet #5010). Glucose dysmetabolism: Fasting glucose and insulin and glucose tolerance testing were quantified after an overnight fast. Liver: Hepatic triglycerides were quantified using Triglyceride Reagent and Triglyceride Standards (Pointe Scientific); serum alanine transaminase (ALT) levels were quantified using ALT Reagent and Catatrol I and II (Catachem); lipid peroxidation was quantified using 4-hydroxynonenal (4-HNE) enzyme-linked immunosorbent assay (ELISA) reagents (Cell Biolabs)—all according to the manufacturer's instructions.

Harley I.T., Stankiewicz T.E., Giles D.A., Softic S., Flick L.M., Cappelletti M., Sheridan R., Xanthakos S.A., Steinbrecher K.A., Sartor R.B., Kohli R., Karp C.L, & Divanovic S. (2014). IL-17 Signaling Accelerates the Progression of Nonalcoholic Fatty Liver Disease in Mice. Hepatology (Baltimore, Md.), 59(5), 1830-1839.

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Quantifying Lipid and Protein Composition

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Phospholipid, total cholesterol, free cholesterol, and triglyceride content of particles were determined enzymatically using the Phospholipids C Kit (Wako Cat# 997-01801), Triglyceride Reagent (Pointe Scientific Cat# T7532), and the Amplex Red Total and Free Cholesterol Assay kit (Thermo Fisher Scientific Cat# A12216). Cholesteryl ester was determined from the difference between total and free cholesterol multiplied by 1.67 to account for the mass of the acyl chain. Protein was determined by the Markwell Lowry assay (35 (link)).

Melchior J.T., Street S.E., Vaisar T., Hart R., Jerome J., Kuklenyik Z., Clouet-Foraison N., Thornock C., Bedi S., Shah A.S., Segrest J.P., Heinecke J.W, & Davidson W.S. (2021). Apolipoprotein A-I modulates HDL particle size in the absence of apolipoprotein A-II. Journal of Lipid Research, 62, 100099.

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Colorimetric Liver Triglyceride Quantification

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Liver triglycerides were measured in ∼100 mg of tissue by colorimetric reactions using GPO (glycerol phosphate oxidase) triglyceride reagent (#T7532-500; Pointe Scientific).

Bugler-Lamb A.R., Hasib A., Weng X., Hennayake C.K., Lin C., McCrimmon R.J., Stimson R.H., Ashford M.L., Wasserman D.H, & Kang L. (2021). Adipocyte integrin-linked kinase plays a key role in the development of diet-induced adipose insulin resistance in male mice. Molecular Metabolism, 49, 101197.

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Liver Triglyceride Extraction and Quantification

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Triglycerides were extracted based on previously established methods (Folch et al., 1957 (link)) with some minor modifications. Approximately 20 – 30 mg of finely powdered liver tissue was added to an ice-cold 10 mL plastic tube and placed on dry ice. Four mL of chloroform/methanol (2:1) was added to the tube and lipids were extracted for 2 h while spinning on a roller. After incubation, 2 mL of 0.6 M NaCl was added and the tubes were briefly vortexed and centrifuged at 2,000 RPM for 10 min at RT. The bottom layer was carefully transferred to a fresh 7 mL glass vial and dried overnight. The samples were resuspended in 500 μL of RNA grade ethanol and vortexed for approximately 5 s. Liver triglyceride concentrations were determined by ELISA using serial dilutions of Precimat Glycerol (Roche; 2.29 nmol/L) as standards. Five ul of standards and samples were added to a 96-well plate and the plate heated for 20 min at 37 °C followed by addition of 300 μL of warm Triglyceride reagent (Pointe Scientific, Inc; Cat #1730711) to each well. The plate was incubated for 30 min at 37 °C and absorbance was measured at 490 nm using a plate reader. Data are expressed as nmol/mg of tissue.

Bernier M., Mitchell S.J., Wahl D., Diaz A., Singh A., Seo W., Wang M., Ali A., Kaiser T., Price N.L., Aon M.A., Kim E.Y., Petr M.A., Cai H., Warren A., Germanio C.D., Di Francesco A., Fishbein K., Guiterrez V., Harney D., Koay Y.C., Mach J., Enamorado I.N., Pulpitel T., Wang Y., Zhang J., Zhang L., Spencer R.G., Becker K.G., Egan J.M., Lakatta E.G., O’Sullivan J., Larance M., LeCouteur D.G., Cogger V.C., Gao B., Fernandez-Hernando C., Cuervo A.M, & de Cabo R. (2020). Disulfiram treatment normalizes body weight in obese mice. Cell metabolism, 32(2), 203-214.e4.

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Serum Lipid and Metabolite Quantification

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TG quantification was performed as reported.14, 17, 18, 19 Briefly, 200 μL of triglyceride reagent (Pointe Scientific) was added to 10 μL of serum in a 96‐well, clear, flat‐bottom plate (Costar). Standards (Pointe Scientific) were prepared according to the manufacturer's instructions. Serum leptin (Millipore) and NO₂ (Sigma‐Aldrich) levels were quantified using commercially available enzyme‐linked immunosorbent assay kits.

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Hepatic Triglyceride Quantification in Mice

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A pre-weighed quantity of frozen mouse liver tissue was placed in 10 μL of Homogenization Buffer (50 mM Tris, 150 mM NaCl, 1 mM EDTA, 1 mM PMSF) per mg of liver tissue. A 1/8” steel bead was added to each sample, and samples were next dissociated using a TissueLyser (Qiagen) for 3 minutes at 30 Hz. Following 20 minutes of incubation on ice, each sample was diluted 1:10 in homogenization buffer and added to a 96 well clear flat bottom plate (Costar) containing 200 μL of Triglyceride Reagent (Pointe Scientific). Standards (Pointe Scientific) were prepared according to manufacturer’s instructions. Hepatic triglycerides were quantified at 500–520 nm using Molecule Devices vmax and Kinetic Microplate Reader and SoftMax Pro v5 software.

Giles D.A., Moreno-Fernandez M.E., Stankiewicz T.E., Cappelletti M., Huppert S.S., Iwakura Y., Dong C., Shanmukhappa S.K, & Divanovic S. (2016). Regulation of Inflammation by IL-17A and IL-17F Modulates Non-Alcoholic Fatty Liver Disease Pathogenesis. PLoS ONE, 11(2), e0149783.

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