75 mg of intestinal tissue was minced, homogenized, and incubated in lysis buffer (50 mM HEPES, pH 7.4, 0.1 % IGEPAL, 150 mM NaCI, 0.25 M sucrose, 0.5 uM AEBSF, and an EDTA free protease inhibitor tablet) on ice for 20 minutes. Insoluble debris was removed by centrifugation at 700 × g for 15 minutes at 4 °C. Protein concentration of the lysate was determined by BCA method, and lysates were equalized with lysis buffer to 6 mg/ml protein concentration for TG quantitation. Triglyceride was quantified using an ‘Infinity TM Triglycerides’ kit (Thermo Scientific, Grand Island, NY) according to manufacture instructions.
Infinity triglycerides kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Infinity Triglycerides kit is a laboratory diagnostic product designed to measure the concentration of triglycerides in biological samples. It provides a quantitative assessment of triglycerides, a type of fat found in the bloodstream.
Automatically generated - may contain errors
Lab products found in correlation
Nefa kit, by Fujifilm (2 mentions)
Mouse ultrasensitive insulin elisa kit, by Crystal Chem (2 mentions)
Aspartate aminotransferase activity kit, by Abcam (2 mentions)
Alanine aminotransferase activity kit, by Abcam (2 mentions)
Mouse leptin elisa, by Crystal Chem (2 mentions)
Microvette cb300 lh k2e tubes, by Sarstedt (1 mentions)
Cholesterol e kit, by Fujifilm (1 mentions)
Vista 1500, by Siemens (1 mentions)
Free fatty acid quantification kit, by Merck Group (1 mentions)
Tg colorimetric assay kit, by Cayman Chemical (1 mentions)
Onetouch ultra, by LifeScan (1 mentions)
Ab156034, by Abcam (1 mentions)
Halt protease phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
15 protocols using infinity triglycerides kit
1
Triglyceride Quantification in Intestinal Tissue
2
Tissue Triglyceride Enzymatic Analysis
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For tissue TG enzymatic analyses, lipids were extracted from tissue homogenates and dissolved in chloroform. The concentrations of TG were measured using a triglyceride assay kit (Infinity TM triglycerides kit, Thermo Fisher Scientific) and normalized to tissue weights as previously described 13 .
3
Hepatic Triglyceride Measurement in Mice
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For the measurement of liver triglyceride contents, liver tissues from Adipo-Cul2-KO or Adipo-Appbp2-KO mice were collected and homogenized in 350 ml ethanolic KOH (100% ethanol and 30% KOH at a ratio of 2:1) and incubated overnight at 55 °C. Subsequently, tissue lysates were supplemented with 50% ethanol to 1 ml final volume. After centrifugation, the supernatant was mixed with 1 M MgCl2 and incubated on ice for 10 min. The amounts of triglycerides were measured using the Infinity Triglycerides kit (Thermo Fisher Scientific). Serum cholesterol and serum triglyceride measurement were performed by the Longwood Small Animal Imaging Facility at BIDMC.
4
Triglyceride Quantification in Liver Tissue
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Frozen liver tissue (50–100 mg) was homogenized in 1 ml PBS. Then, 800 μl lysates were added to 4 ml extraction buffer. After thoroughly rotating for 30 min at room temperature (RT), the lipid phase was separated from the aqueous phase by centrifuging at 1,800g for 20 min. A 0.2-ml lipid fraction in the organic phase was collected and transferred to a 1.5-ml tube to dry under a nitrogen stream in the fume hood. Then, 0.2 ml 2% Triton X-100 solution was used to solubilize the lipids. TG levels were determined using the Infinity Triglycerides kit (Thermo Fisher). The lipid amount was normalized to the liver lysate protein amount.
5
Plasma NEFA and TG Quantification
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Blood was collected from the tail vein of mice in Microvette CB300 LH K2E tubes (Sarstedt). Plasma NEFA and TG levels were measured by colorimetrical analyses using the WAKO NEFA analysis kit (NEFA-HR(2); Wako Pure Chemical Industries) and the Infinity™ triglycerides kit, (Thermo Fisher Scientific), respectively.
6
Plasma Metabolic Biomarker Analysis
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Whole blood was taken from the facial vein and blood glucose was measured with a glucose meter (Easy Touch) from the tail vein. Plasma was collected after centrifugation at 1,200 x rpm, 4°C for 10 min. Plasma triglyceride (TG) and free fatty acid (FFA) levels were measured with Infinity™ Triglycerides kit (Thermo Fisher) and NEFA kit (WAKO). Plasma insulin levels were measured with the Mouse Ultrasensitive Insulin ELISA kit (Crystal Chem, #90080), and leptin levels were measured with Mouse Leptin ELISA (Crystal Chem, #90030) kit. Plasma AST and ALT activity was measured with the Aspartate Aminotransferase Activity kit (Biovision, #K753) and Alanine Aminotransferase Activity kit (Biovision, #K752).
7
Metabolic Biomarkers in Experimental Analyses
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Serum total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C), triglycerides (TGs), glucose (Glu), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels were measured by the enzymatic procedures run on Siemens Vista 1500 instrumentation (Clinical laboratory of Richmond VA Medical Center). Serum insulin was measured by the sensitive ELISA sandwich assay method using Crystal Chem Ultra-sensitive mouse insulin ELISA kit (Elk Grove Village, IL, catalogue # 90080) according to manufacturer’s instruction. IR was assessed by using homeostasis model assessment of insulin resistance (HOMA-IR) score calculated as [glucose (mg/dl) x insulin (ng/ml) × 26 / 405]. Liver TC was measured by the enzymatic assay using the Cholesterol E Kit (FUJIFILM Wako Chemicals, catalogue # 999-02601). Infinity Triglycerides Kit (Thermo Fisher Scientific, catalogue # 22421) was used for liver TG measurement. Abcam free fatty acid (FFA) assay kit (catalogue # Ab65341) was used for the liver FFA measurements.
8
Liver Lipid Extraction and Quantification
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Snap frozen liver tissues were homogenized in 1.5 ml Chloroform/methanol (2:1 v/v). Lipids were measured using Infinity Triglycerides kit (Thermo Fischer Scientific).
9
Lipid Profile Determination Protocol
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Plasma levels of cholesterol and triglyceridess were determined by enzymatic means using a chemistry autoanalyzer (CX7; Beckman Coulter Diagnostics, Fullerton, California, USA). Plasma levels of triglycerides were measured with an Infinity Triglycerides kit (Thermo Scientific, Waltham, Massachusetts, USA). Levels of high-density lipoprotein-cholesterol (HDL-C) and low-density lipoprotein-cholesterol (LDL-C) were determined using homogeneous assay kits (Equal Diagnostics, Exton, Pennsylvania, USA).
10
Metabolic Biomarkers in Animal Model
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Blood glucose was measured by a blood glucose monitor (OneTouch Ultra; LifeScan, Inc., Milpitas, CA, USA). Plasma insulin, leptin, and corticosterone concentrations were determined by EIA using commercially available assay kits (ALPCO Diagnostics, Salem, NH): insulin ELISA (80-INSMR), leptin ELISA (22-LEPMS), and corticosterone ELISA (55-CORMS). Belfiore insulin sensitivity index (ISI) (pmol/L*mmol/L*h) was calculated based on the formula: 2/((insulin GTT AUC * glucose GTT AUC) + 1). HOMA-IR was calculated based on the formula: fasting glucose (mmol/L) × fasting insulin (mU/L)/22.5. Hepatic triglyceride was extracted from liver tissue that was homogenized in 5% NP-40, heated to 90°C for 3 min, centrifuged at 10,000 g for 10 min to collect the supernatant. The triglyceride content in the supernatant was subsequently determined by TG Colorimetric Assay Kit (Item No. 10010303; Cayman Chemical, Ann Arbor, MI, USA). Plasma triglycerides were measured with Infinity™ Triglycerides Kit (TR22421; Thermo Fisher Scientific). Plasma free fatty acids (FFAs) were measured using a Free Fatty Acid Quantification Kit (MAK044; Sigma).
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