Blood glucose was measured with a Glucometer – Contour XT (Bayer). Plasma Insulin and Adiponectin were measured with ELISAs from Alpco (Alpco Salem, BioCatGmbH). Plasma HSP60 was measured with an ELISA from Elabscience (Elabscience Inc.). Leptin was measured using an ELISA from R & D Systems (R & D Systems/Bio-Techne GmbH) and Plasma Glucagon levels with an ELISA from Yanaihara (Yanaihara Institute Inc.). FGF-21 was measured using an ELISA from Biovendor (BioVendor R&D). All ELISAs were performed according to the manufacturer's guidelines. Triacylglycerols were measured with Triglyceride Reagent ABX, and NEFAs were measured with the NEFA-HR assay according to manufacturer's guidelines (FUJIFILM Wako Chemicals Europe GmbH). The glucose in the supernatants of cultured 3T3-L1 cells was determined by the hexokinase method (Roche Diagnostics GmbH).
Nefa hr assay
Manufactured by Fujifilm
Sourced in United States, Germany
The NEFA-HR assay is a laboratory equipment product developed by Fujifilm. It is designed to measure the concentration of non-esterified fatty acids (NEFAs) in biological samples. The NEFA-HR assay provides a reliable and efficient method for analyzing NEFA levels, which can be useful in various clinical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Contour xt, by Bayer (3 mentions)
Absciex 6500 qtrap, by Shimadzu (1 mentions)
Serum triglyceride determination kit, by Merck Group (1 mentions)
Glycogen assay kit 2, by Abcam (1 mentions)
Ultra sensitive mouse insulin elisa kit, by Crystal Chem (1 mentions)
Corticosterone elisa kit, by Abcam (1 mentions)
Leptin, by R&D Systems (1 mentions)
4 protocols using nefa hr assay
1
Comprehensive Metabolic Biomarker Analysis
2
Metabolomic Analysis of Cell Cultures
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We measured glucose and lactate concentrations enzymatically using the YSI 2500 Glucose/Lactate Analyzer (Yellow Springs, OH, USA). We measured glutamine concentration and enrichment by LC-MS/MS (AbSCIEX 6500 QTRAP with a Shimadzu ultrafast liquid chromatography system in negative ion mode) as we have previously reported [25 (link)] after spiking with [2H4] taurine as an internal standard. We measured citrate, succinate, and malate concentration and enrichment using the same LC-MS/MS method. Mass/charge ratios for the unlabeled metabolites are shown in Table 2 . Due to space constraints, the labeled intermediate mass/charge ratios are not shown.
Mass/charge ratios for the unlabeled metabolites
| Analyte | Mass/charge |
|---|---|
| Citrate | 191/173 |
| Glutamate | 146/128 |
| Malate | 133/115 |
| Succinate | 117/99 |
3
Measuring Metabolic Biomarkers in Exercise
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Blood was collected via tail vein immediately following the exercise bout. Blood glucose concentration was measured via a hand-held glucometer (Contour XT, Bayer). Serum non-esterified fatty acids (NEFA) were assessed using the NEFA-HR Assay (FujiFilm). Serum Catecholamines (Adrenaline and Noradrenaline) were assessed using a 2-CAT Research ELISA (LDN). Serum Corticosterone was assessed using a Corticosterone ELISA kit (ab108821) (Abcam). Serum triglycerides were determined using a Serum Triglyceride Determination Kit (Sigma Aldrich). Serum insulin was determined using an Ultra-Sensitive Mouse Insulin ELISA Kit (Crystal Chem). Liver and gastrocnemius glycogen content were assessed using a Glycogen Assay Kit II (Abcam).
4
Glucose, Insulin, and Lipid Analysis
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Blood glucose was analyzed with a glucometer (Contour XT, Bayer AG, Leverkusen, Germany). ELISA kits from the following manufactures were used: Plasma and pancreatic Insulin Alpco (Alpco, Salem, MA, USA); Leptin, R&D Systems (R&D Systems/Bio-Techne GmbH, Wiesbaden, Germany). The manufacturer’s recommendations were followed. Triacylglycerols (TAGs) were measured with Triglyceride Reagent from ABX. Non-esterified fatty acids (NEFA) were measured with NEFA-HR Assay (FUJIFILM Wako Chemicals Europe GmbH, Neuss, Germany).
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