Infinity triglycerides reagent

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Infinity Triglycerides Reagent is a laboratory product designed for the quantitative determination of triglycerides in biological samples. It provides a reliable and accurate method for measuring triglyceride levels.

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Lab products found in correlation

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56 protocols using infinity triglycerides reagent

Muscle Triglyceride Quantitation Protocol

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Muscle triglyceride content was measured by the modified Bligh and Dyer method [23 (link)]. Approximately 60 mg skeletal muscle tissue was homogenized in 1 mL of ice-cold methanol and lipids were extracted using 1:2 methanol:chloroform. The polar and non-polar phases were then separated by centrifugation, and the lower non-polar phase was isolated and dried under nitrogen. The dried samples were then resuspended in isopropanol + 2% Triton X-100 for direct triglyceride quantitation using Infinity Triglycerides Reagent (Thermo Scientific, Waltham, MA, USA) and a standard curve with glycerol standard solution (Sigma-Aldrich, St. Louis, MO, USA) per manufacturer’s instructions. Resultant triglyceride concentrations were normalized to starting tissue weight. Results from dams and select offspring measures have been reported previously, as were fetal and adult liver triglyceride content [17 (link)].

Boyle K.E., Magill-Collins M.J., Newsom S.A., Janssen R.C, & Friedman J.E. (2020). Maternal Fat-1 Transgene Protects Offspring from Excess Weight Gain, Oxidative Stress, and Reduced Fatty Acid Oxidation in Response to High-Fat Diet. Nutrients, 12(3), 767.

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Quantifying Hepatic Triglyceride Levels

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Triglyceride content within mouse liver was analyzed as previously described [67 (link)]. In brief, liver was homogenized in NETN lysis buffer supplemented with 5 mM NaF (Sigma-Aldrich, St. Louis, MO), 50 mM 2-glycerophosphate (Sigma-Aldrich), and protease inhibitor cocktail (Roche, Nutley, NJ). Lipid content was determined using Infinity Triglycerides Reagent (Thermo Fisher Scientific, Waltham, MA) according to the manufacturer's protocol.

Stout M.B., Tchkonia T., Pirtskhalava T., Palmer A.K., List E.O., Berryman D.E., Lubbers E.R., Escande C., Spong A., Masternak M.M., Oberg A.L., LeBrasseur N.K., Miller R.A., Kopchick J.J., Bartke A, & Kirkland J.L. (2014). Growth hormone action predicts age-related white adipose tissue dysfunction and senescent cell burden in mice. Aging (Albany NY), 6(7), 575-586.

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Triglyceride Quantification in Cultured Cells

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Triglyceride (TG) levels were determined using a commercial kit (Infinity™ Triglycerides Reagent, Thermo Fisher Scientific, Middleton, VA, USA). Briefly, after 48 h, cells were washed twice with phosphate-buffered saline (PBS) and harvested by scraping the cells from the culture plate in PBS containing 1 % Triton-X. Cell homogenates were obtained by sonication using Fisherbrand™ Model 50 Sonic Dismembrator (Fisher Scientific), and TG concentrations were determined according to the manufacturer’s instructions. Protein concentrations were measured using Pierce BCA Protein Assay Kit and used for normalization of samples.

Qi W., Clark J.M., Timme-Laragy A.R, & Park Y. (2020). Perfluorobutanesulfonic Acid (PFBS) Induces Fat Accumulation in HepG2 Human Hepatoma. Toxicological and environmental chemistry, 102(10), 585-606.

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Hepatic Lipid Extraction and Quantification

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As described previously [14 (link)], hepatic lipids were extracted and saponified by the Folch extraction method. After saponification, TG were measured spectrophotometrically using the Infinity Triglycerides reagent (Thermo Fisher Scientific (Middletown, VA, USA). A NEFA-HR (2) diagnostic kit from Wako Life Sciences, Inc. (Mountain View, CA, USA).was used to measure serum NEFA.

Rasineni K., Kubik J.L., Casey C.A, & Kharbanda K.K. (2019). Inhibition of Ghrelin Activity by Receptor Antagonist [d-Lys-3] GHRP-6 Attenuates Alcohol-Induced Hepatic Steatosis by Regulating Hepatic Lipid Metabolism. Biomolecules, 9(10), 517.

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Quantitative Lipid Analysis of Liver

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Liver tissues were washed in 50 mM NaCl and homogenized in 1 mL 50 mM NaCl with 0.5 mm glass silica beads and beaded for 30 s. Hepatic lipids were extracted using an aqueous solution of chloroform and methanol in a 2:1 ratio based on the Bligh and Dyer method (Bligh and Dyer, 1959). Triglyceride and cholesterol standards (catalog: T7531-STD, C7509-STD; Point Scientific; Canton, MI) were utilized to generate a standard curve to quantify the extracted lipids. Extracted triglycerides and cholesterol were colorimetrically quantified using a microplate absorbance reader (BioTek Gen 5; Winooski, VT, USA). Reagents used in the assay: Infinity Triglycerides Reagent (TR22421, ThermoFisher Scientific, Middletown, VA, USA) and Infinity Cholesterol Reagent (TR13421, ThermoFisher Scientific, Middletown, VA, USA).

Bolatimi O.E., Head K.Z., Luo J., Gripshover T.C., Lin Q., Adiele N.V., Watson W.H., Wilkerson C., Cai L., Cave M.C, & Young J.L. (2023). Can Zinc Supplementation Attenuate High Fat Diet-Induced Non-Alcoholic Fatty Liver Disease?. International Journal of Molecular Sciences, 24(2), 1763.

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Generation and Characterization of A1CF Knock-in Mice

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All procedures used for animal studies were approved by Harvard University’s Faculty of Arts and Sciences Institutional Animal Care and Use Committee and were consistent with local, state, and federal regulations as applicable. Knock-in mice were generated using a guide RNA designed to target the orthologous site of the A1CF p.Gly390Ser variant. In vitro transcribed Cas9 mRNA (100 ng/μL; TriLink BioTechnologies) and guide RNA (50 ng/μL) were co-injected with 100 ng/μL single-strand DNA oligonucleotide (Integrated DNA Technologies): (Supplementary Table 21) into the cytoplasm of fertilized oocytes from C57BL/6J mice. Genomic DNA samples from founder mice were screened for knock-in mutations by PCR and confirmed by Sanger sequencing. Positive mice were bred with C57BL/6J mice to generate wild-type and homozygous knock-in mice. Male colony mates at 12 weeks of age were used for lipid measurements. Blood samples were collected from the lateral tail vein following an overnight fast. Plasma triglyceride levels were measured using Infinity Triglycerides Reagent (Thermo Fisher) according to the manufacturers’ instructions.

Liu D.J., Peloso G.M., Yu H., Butterworth A.S., Wang X., Mahajan A., Saleheen D., Emdin C., Alam D., Alves A.C., Amouyel P., di Angelantonio E., Arveiler D., Assimes T.L., Auer P.L., Baber U., Ballantyne C.M., Bang L.E., Benn M., Bis J.C., Boehnke M., Boerwinkle E., Bork-Jensen J., Bottinger E.P., Brandslund I., Brown M., Busonero F., Caulfield M.J., Chambers J.C., Chasman D.I., Chen Y.E., Chen Y.D., Chowdhury R., Christensen C., Chu A.Y., Connell J.M., Cucca F., Cupples L.A., Damrauer S.M., Davies G., Deary I.J., Dedoussis G., Denny J.C., Dominiczak A., Dubé M.P., Ebeling T., Eiriksdottir G., Esko T., Farmaki A.E., Feitosa M.F., Ferrario M., Ferrieres J., Ford I., Fornage M., Franks P.W., Frayling T.M., Frikke-Schmidt R., Fritsche L., Frossard P., Fuster V., Ganesh S.K., Gao W., Garcia M.E., Gieger C., Giulianini F., Goodarzi M.O., Grallert H., Grarup N., Groop L., Grove M.L., Gudnason V., Hansen T., Harris T.B., Hayward C., Hirschhorn J.N., Holmen O.L., Huffman J., Huo Y., Hveem K., Jabeen S., Jackson A.U., Jakobsdottir J., Jarvelin M.R., Jensen G.B., Jørgensen M.E., Jukema J.W., Justesen J.M., Kamstrup P.R., Kanoni S., Karpe F., Kee F., Khera A.V., Klarin D., Koistinen H.A., Kooner J.S., Kooperberg C., Kuulasmaa K., Kuusisto J., Laakso M., Lakka T., Langenberg C., Langsted A., Launer L.J., Lauritzen T., Liewald D.C., Lin L.A., Linneberg A., Loos R.J., Lu Y., Lu X., Mägi R., Malarstig A., Manichaikul A., Manning A.K., Mäntyselkä P., Marouli E., Masca N.G., Maschio A., Meigs J.B., Melander O., Metspalu A., Morris A.P., Morrison A.C., Mulas A., Müller-Nurasyid M., Munroe P.B., Neville M.J., Nielsen J.B., Nielsen S.F., Nordestgaard B.G., Ordovas J.M., Mehran R., O’Donnell C.J., Orho-Melander M., Molony C.M., Muntendam P., Padmanabhan S., Palmer C.N., Pasko D., Patel A.P., Pedersen O., Perola M., Peters A., Pisinger C., Pistis G., Polasek O., Poulter N., Psaty B.M., Rader D.J., Rasheed A., Rauramaa R., Reilly D., Reiner A.P., Renström F., Rich S.S., Ridker P.M., Rioux J.D., Robertson N.R., Roden D.M., Rotter J.I., Rudan I., Salomaa V., Samani N.J., Sanna S., Sattar N., Schmidt E.M., Scott R.A., Sever P., Sevilla R.S., Shaffer C.M., Sim X., Sivapalaratnam S., Small K.S., Smith A.V., Smith B.H., Somayajula S., Southam L., Spector T.D., Speliotes E.K., Starr J.M., Stirrups K.E., Stitziel N., Strauch K., Stringham H.M., Surendran P., Tada H., Tall A.R., Tang H., Tardif J.C., Taylor K.D., Trompet S., Tsao P.S., Tuomilehto J., Tybjaerg-Hansen A., van Zuydam N.R., Varbo A., Varga T.V., Virtamo J., Waldenberger M., Wang N., Wareham N.J., Warren H.R., Weeke P.E., Weinstock J., Wessel J., Wilson J.G., Wilson P.W., Xu M., Yaghootkar H., Young R., Zeggini E., Zhang H., Zheng N.S., Zhang W., Zhang Y., Zhou W., Zhou Y., Zoledziewska M., Howson J.M., Danesh J., McCarthy M.I., Cowan C., Abecasis G., Deloukas P., Musunuru K., Willer C.J, & Kathiresan S. (2017). Exome-wide association study of plasma lipids in >300,000 individuals. Nature genetics, 49(12), 1758-1766.

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Triglyceride Content Quantification in Adipocytes

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Following cell treatment, the medium was removed, and mature adipocytes were thoroughly washed with phosphate-buffered saline (PBS) and harvested using a 300 µL of buffer comprising Tris-HCl at pH 7.4, 0.15 M sodium chloride (NaCl) and 1 mM ethylene diamine tetra acetate (EDTA) with protease inhibitors (0.1 M phenylmethylsulphonyl fluoride and 0.1 M iodoacetamide). Afterwards, cell samples were disrupted using ultrasound equipment, the Branson Digital Sonifier SFX 550 (Emerson Electric Co, St. Louis, MO, USA) with a 2 mm diameter ultrasound-microprobe (Biogen Scientific S.L., Madrid, Spain). Triglyceride content (mg/mL) was measured using Infinity Triglycerides reagent (Thermo Scientific, Rockford, IL, USA). The lipid content of cells was standardised by the protein content of each well. Protein content was determined using the method described by Bradford et al. [33 (link)]. Results are expressed as mg of triglycerides/mg of protein, and presented in arbitrary units.

Gómez-López I., Eseberri I., Cano M.P, & Portillo M.P. (2024). Anti-Obesity Effect of Different Opuntia stricta var. dillenii’s Prickly Pear Tissues and Industrial By-Product Extracts in 3T3-L1 Mature Adipocytes. Nutrients, 16(4), 499.

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Serum Lipid and Glucose Quantification

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Serum NEFA, TG, and cholesterol concentrations were determined by using commercial kits for NEFA (Wako Diagnostics, VA), TG (Infinity Triglycerides Reagent, Thermo Scientific, USA), and cholesterol (Infinity total cholesterol, Thermo Scientific, USA). These assays were performed by the Diabetes Research Center (DRC) at Washington University School of Medicine. Blood glucose concentrations were measured by using commercial blood test strips (Accu-Check, Roche, USA) by using a drop of tail blood.

Devanathan S., Whitehead T., Schweitzer G.G., Fettig N., Kovacs A., Korach K.S., Finck B.N, & Shoghi K.I. (2014). An Animal Model with a Cardiomyocyte-Specific Deletion of Estrogen Receptor Alpha: Functional, Metabolic, and Differential Network Analysis. PLoS ONE, 9(7), e101900.

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Lipid Extraction and Quantification

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Quadriceps and 100–200 mg liver samples were collected from overnight‐fasted mice, flash frozen in liquid nitrogen, and stored at −80°C until analysis. Frozen tissues were homogenized in 7.5 mL chloroform/methanol (2:1), sonicated for 1 h, and filtered using syringe‐driven 0.45 μm PTFE filter units. Sulfuric acid (2.5 mL of 0.05%) was added to filtered extractions and samples were centrifuged for 2 min at 2500 g. The organic phase was allowed to evaporate for 48 h and then reconstituted in 100% chloroform. Duplicate 500 μL aliquots were dried overnight to determine total lipids. Additional aliquots were used to determine triglyceride content using Infinity^™ Triglycerides Reagent (Thermo Fisher Scientific, Waltham, MA) and cholesterol levels using an enzymatic kit (Wako Diagnostics, Richmond, VA).

Hammond C.L., Wheeler S.G., Ballatori N, & Hinkle P.M. (2015). Ostα−/− mice are not protected from western diet‐induced weight gain. Physiological Reports, 3(1), e12263.

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Plasma and Hepatic Lipid Profiling

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Plasma levels for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and circulating levels of high-density lipoprotein (HDL), triglycerides, and cholesterol were obtained using an Abaxis Piccolo Xpress analyzer (Abbott; Abbott Park, IL, USA) with Lipid Panel Plus diskettes (catalog:400-0030; Abaxis Inc.; Union City, CA, USA). Circulating levels of plasma adipokines and cytokines were measured using a MILLIPLEX^® Mouse Adipokine Magnetic Bead Panel (Millipore Sigma, Burlington, MA, USA) for analytes interleukin 6 (IL-6), leptin, total plasminogen activator inhibitor 1 (PAI-1), tumor necrosis factor alpha (TNFα), and resistin. Data was obtained from a Luminex 100/200 System equipped with xPONENT Software (Luminex Corporate, Austin, TX, USA). Hepatic lipids were extracted using a modified Bligh and Dyer method with a 2:1 ratio of chloroform to methanol mixture (27 (link), 28 (link)). Extracted lipids were measured using Infinity Assay Reagents: Infinity Triglycerides reagent (TR22421, ThermoFisher Scientific, Middletown, VA, USA) and Infinity Cholesterol reagent (TR13421, ThermoFisher Scientific, Middletown, VA, USA). Sample absorbance at 500 nm was calculated from a standard curve constructed from clinical standards (catalog: T7531-STD, C7509-STD; Point Scientific; Canton, MI) for each analyte ran simultaneously with unknown samples.

Griscelli F., Li H., Bennaceur-Griscelli A., Soria J., Opolon P., Soria C., Perricaudet M., Yeh P, & Lu H. (1998). Angiostatin gene transfer: Inhibition of tumor growth in vivo by blockage of endothelial cell proliferation associated with a mitosis arrest. Proceedings of the National Academy of Sciences of the United States of America, 95(11), 6367-6372.

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