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Infinity tg reagent

Manufactured by Thermo Fisher Scientific
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Sourced in Germany

The Infinity TG reagent is a laboratory diagnostic product designed for the quantitative determination of triglycerides in human serum or plasma. It utilizes an enzymatic colorimetric method to measure the triglyceride concentration.

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Lab products found in correlation

Trehalase solution, by Merck Group (2 mentions) Glucose hk assay kit, by Merck Group (2 mentions) Hr series nefa hr 2 kit, by Fujifilm (2 mentions) Spectramax 250 microplate reader, by Molecular Devices (2 mentions) Xevo tq ms acquity uplc system, by Waters Corporation (2 mentions) Nefa c, by Fujifilm (2 mentions) Free glycerol reagent, by Merck Group (2 mentions) Infinity cholesterol reagent, by Thermo Fisher Scientific (1 mentions) Ffa quantification kit, by Abcam (1 mentions) Onetouch ultra 2 blood glucose meter, by LifeScan (1 mentions) Cholesterol assay kit, by Abcam (1 mentions) Accutrend plus, by Roche (1 mentions) Mouse leptin elisa, by Crystal Chem (1 mentions)

15 protocols using infinity tg reagent

1

Measuring Fly Triglycerides and Trehalose

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For TG assay, flies were homogenized in 300 µl of 0.05% PBS-Triton, and the supernatant was heated for 10 min at 70°C to inactivate endogenous enzymes. 300 µl of Infinity TG Reagent (Thermo Electron Corp) was added to 20 µl homogenate and incubated at 37°C for 15 min. TG levels were measured as the OD value at 520 nm in Spectra Max 340 Microplate Reader and total amounts were determined using TG standard. For trehalose assay, flies were homogenized in a buffer containing 5 mM Tris pH 6.6, 137 mM NaCl and 2.7 mM KCl, followed by a heat treatment for 10 min at 70 °C to inactivate endogenous trehalase. Half of the supernatant was diluted with an equal volume of the buffer to provide the basal value, the other half was diluted with trehalase solution (Sigma) to break trehalose into free glucose at 37 °C for 20 hours, and glucose levels were measured using Glucose (HK) Assay Kit (sigma) via reading of OD value at 340 nm. To measure the circulating trehalose, hemolymph was extracted by decapitation and centrifugation from 30–50 adult flies, and an aliquot of collected hemolymph was used to measure trehalose using the assay described above.
Zhang Y., Liu G., Yan J., Zhang Y., Li B, & Cai D. (2015). Metabolic learning and memory formation by the brain influence systemic metabolic homeostasis. Nature communications, 6, 6704.
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2

Measuring Fly Triglycerides and Trehalose

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For TG assay, flies were homogenized in 300 µl of 0.05% PBS-Triton, and the supernatant was heated for 10 min at 70°C to inactivate endogenous enzymes. 300 µl of Infinity TG Reagent (Thermo Electron Corp) was added to 20 µl homogenate and incubated at 37°C for 15 min. TG levels were measured as the OD value at 520 nm in Spectra Max 340 Microplate Reader and total amounts were determined using TG standard. For trehalose assay, flies were homogenized in a buffer containing 5 mM Tris pH 6.6, 137 mM NaCl and 2.7 mM KCl, followed by a heat treatment for 10 min at 70 °C to inactivate endogenous trehalase. Half of the supernatant was diluted with an equal volume of the buffer to provide the basal value, the other half was diluted with trehalase solution (Sigma) to break trehalose into free glucose at 37 °C for 20 hours, and glucose levels were measured using Glucose (HK) Assay Kit (sigma) via reading of OD value at 340 nm. To measure the circulating trehalose, hemolymph was extracted by decapitation and centrifugation from 30–50 adult flies, and an aliquot of collected hemolymph was used to measure trehalose using the assay described above.
Zhang Y., Liu G., Yan J., Zhang Y., Li B, & Cai D. (2015). Metabolic learning and memory formation by the brain influence systemic metabolic homeostasis. Nature communications, 6, 6704.
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3

Quantification of Circulating and Hepatic Lipids

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The concentration of circulating free fatty acids (FFAs) and TGs were directly measured in plasma as described below. The concentration of hepatic FFAs and TGs were measured in lipid extracts obtained using a Folch solution (chloroform and methanol; 2:1 ratio; (Folch et al., 1957 (link))). Lipid extracts were solubilized in a 2% Triton X-100 solution prior to analysis. The concentration of FFAs in samples was determined using a HR Series NEFA-HR(2) kit according to the manufacturer’s instructions (Wako Diagnostics, Richmond, VA). The concentration of TG in samples was determined using an Infinity TG reagent according to the manufacturer’s instructions (Thermo Fisher Scientific, Waltham, MA). All protocols were adapted to a 96-well plate format while preserving sample-to-reagent ratios. Plates were read using a SpectraMax 250 Microplate Reader (Molecular Devices, Sunnyvale, CA). The individual acyl composition of plasma and liver FFA was obtained by LC/MS using a Waters Xevo TQ MS ACQUITY UPLC system (Waters, Milford, MA), according to our previously published method (Clugston et al., 2011 (link)).
Huang L., Yuen J.J., Trites M.J., Saha A., Epps C.T., Hu Y., Kerolle S., Lee S., Jiang H., Goldberg I.J., Blaner W.S, & Clugston R.D. (2018). Dietary Macronutrient Composition Determines the Contribution of DGAT1 to Alcoholic Steatosis. Alcoholism, Clinical and Experimental Research, 42(12), 2298-2312.
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4

Blood Lipid and Glucose Measurement

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Blood glucose was measured using a One-Touch Ultra2 blood glucose meter (Life Scan, Milpitas, CA). Plasma TG and cholesterol concentrations were measured with the Infinity TG Reagent and Infinity Cholesterol Reagent (Thermo Scientific, Waltham, MA), respectively. Concentrations of plasma free fatty acids (FFA) were determined with a FFA Quantification Kit (BioVision, Mountain View, CA).
Zhang W., Sun Q., Zhong W., Sun X, & Zhou Z. (2016). Hepatic peroxisome proliferator-activated receptor gamma signaling contributes to alcohol-induced hepatic steatosis and inflammation in mice. Alcoholism, clinical and experimental research, 40(5), 988-999.
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5

Quantifying Lipid and Cholesterol Levels

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Total lipids were extracted with modified isopropanol:hexane extraction method.(16) TG content in the dried lipids was quantified using the Infinity TG reagent (Thermo Fisher Scientific). For cholesterol, lipids were extracted from liver homogenates with chloroform:isopropanol:NP‐40 (7:11:0.1). The dried lipids were dissolved with assay buffer. The levels of total cholesterol were measured using a cholesterol assay kit according to the manufacturer’s instructions (Abcam, Cambridge, MA). The TG and cholesterol levels in cells or liver tissue were expressed as the ratio of total TG or cholesterol content in the cell lysate/tissue homogenate to the total protein levels in the lysate/homogenate.
Athinarayanan S., Fan Y., Wang X., Callaway E., Cai D., Chalasani N., Chapkin R.S, & Liu W. (2020). Fatty Acid Desaturase 1 Influences Hepatic Lipid Homeostasis by Modulating the PPARα‐FGF21 Axis. Hepatology Communications, 5(3), 461-477.
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6

Lipid Extraction and Triglyceride Quantification

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Lipids were extracted from PPDIV cells using a 3:2 mixture of Hexane:Isopropanol. Liver tissue was weighed and homogenized and lyophilized, then lipids were extracted with chloroform. The organic solvent was evaporated from the extracted lipids and triglycerides content was quantified using the infinity TG reagent (Thermo-Fisher) following the manufacturer’s protocol.
Reilly S.M., Hung C.W., Ahmadian M., Zhao P., Keinan O., Gomez A.V., DeLuca J.H., Dadpey B., Lu D., Zaid J., Poirer B., Peng X., Yu R.T., Downes M., Liddle C., Evans R.M., Murphy A.N, & Saltiel A.R. (2020). Catecholamines suppress fatty acid re-esterification and increase oxidation in white adipocytes via STAT3. Nature metabolism, 2(7), 620-634.
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7

VLDL-TG Secretion Rate Measurement

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To measure the VLDL-TG secretion rate, mice were fasted for 6 h, pre bled by retro orbital bleeding, and injected intravenously with 10% tyloxapol (Triton WR-1339; T-8761; Sigma-Aldrich, St. Louis, MO; 500 mg/kg body weight) to inhibit lipolysis. Plasma samples were drawn serially 0, 30, 60, 90 and 120 min after injection. Plasma TG from each time was measured by plate assay using Infinity TG Reagent (Thermo Scientific).
Ding Y., Xian X., Holland W.L., Tsai S, & Herz J. (2016). Low-Density Lipoprotein Receptor-Related Protein-1 Protects Against Hepatic Insulin Resistance and Hepatic Steatosis. EBioMedicine, 7, 135-145.
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8

Quantification of Circulating and Hepatic Lipids

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The concentration of circulating free fatty acids (FFAs) and TGs was directly measured in plasma as described below. The concentration of hepatic FFAs and TGs was measured in lipid extracts obtained using a Folch solution (chloroform and methanol; 2:1 ratio; Folch et al., 1957). Lipid extracts were solubilized in a 2% Triton X‐100 solution prior to analysis. The concentration of FFAs in samples was determined using a HR Series NEFA‐HR(2) kit according to the manufacturer's instructions (Wako Diagnostics, Richmond, VA). The concentration of TG in samples was determined using an Infinity TG reagent according to the manufacturer's instructions (Thermo Fisher Scientific, Waltham, MA). All protocols were adapted to a 96‐well plate format while preserving sample‐to‐reagent ratios. Plates were read using a SpectraMax 250 Microplate Reader (Molecular Devices, Sunnyvale, CA). The individual acyl composition of plasma and liver FFA was obtained by liquid chromatography‐mass spectrometry (LC/MS) using a Waters Xevo TQ MS ACQUITY UPLC system (Waters, Milford, MA), according to our previously published method (Clugston et al., 2011).
Huang L., Yuen J.J., Trites M.J., Saha A., Epps C.T., Hu Y., Kerolle S., Lee S., Jiang H., Goldberg I.J., Blaner W.S, & Clugston R.D. (2018). Dietary Macronutrient Composition Determines the Contribution of DGAT1 to Alcoholic Steatosis. Alcoholism, Clinical and Experimental Research, 42(12), 2298-2312.
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9

Colorimetric Assay for Total Lipids

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Total lipid content was determined using a triglyceride colorimetric assay. Samples were finely cut, then lipids were extracted by the Schwartz method using organic solvents and concentrated by evaporation. Enzymatic hydrolysis of triglycerides by lipase to glycerol and free fatty acids was carried out using Infinity TG Reagent (Thermo Fisher Scientific) according to the manufacturer’s protocol. Glycerol was detected by absorbance at 540 nm. Absolute glycerol concentrations were determined from a standard curve and the percentage of lipids removed from AAT was determined relative to control adipose samples.
Anderson A.E., Wu I., Parrillo A.J., Wolf M.T., Maestas DR J.r., Graham I., Tam A.J., Payne R.M., Aston J., Cooney C.M., Byrne P., Cooney D.S, & Elisseeff J.H. (2022). An immunologically active, adipose-derived extracellular matrix biomaterial for soft tissue reconstruction: concept to clinical trial. NPJ Regenerative Medicine, 7, 6.
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10

Lipid Extraction and Triglyceride Quantification

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Lipids were extracted from PPDIV cells using a 3:2 mixture of Hexane:Isopropanol. Liver tissue was weighed and homogenized and lyophilized, then lipids were extracted with chloroform. The organic solvent was evaporated from the extracted lipids and triglycerides content was quantified using the infinity TG reagent (Thermo-Fisher) following the manufacturer’s protocol.
Reilly S.M., Hung C.W., Ahmadian M., Zhao P., Keinan O., Gomez A.V., DeLuca J.H., Dadpey B., Lu D., Zaid J., Poirer B., Peng X., Yu R.T., Downes M., Liddle C., Evans R.M., Murphy A.N, & Saltiel A.R. (2020). Catecholamines suppress fatty acid re-esterification and increase oxidation in white adipocytes via STAT3. Nature metabolism, 2(7), 620-634.
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