Plasma glucose was measured enzymatically as previously described [22 ]. Plasma lactate was measured from PCA extract of plasma samples in a reaction mix containing glycyl-glycine, H2O, NAD+ and glutamic acid using the same general principles as for plasma glucose. NEFA content was determined using WAKO HR Series NEFA-HR (2) (WAKO Diagnostics, Richmond, VA, USA) according to manufacturer’s guidelines.
Wako hr series nefa hr 2
Manufactured by Fujifilm
Sourced in United States
The WAKO HR Series NEFA-HR (2) is a laboratory instrument used for the quantitative determination of non-esterified fatty acids (NEFA) in biological samples. It functions as a diagnostic tool for measuring NEFA levels, which are important indicators of various metabolic and physiological conditions.
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Lab products found in correlation
4 protocols using wako hr series nefa hr 2
1
Plasma Metabolite Measurement Protocol
2
Fatty Acid Preparation for Assays
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Oleic acid (OA) (Sigma) and palmitic acid (PA) (Sigma) were dissolved in Krebs‐Ringer bicarbonate buffer complexed with 5% fatty‐acid free BSA (Sigma) under gentle heating and stirring and sterile‐filtered through a 0.22 μm filter. FA concentration was quantified using Wako HR series NEFA‐HR(2) according to manufacturer instructions.
3
Blood Lipid Quantification by Enzymatic Assays
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Enzymatic colorimetric assays of Triacylglycerol (TAG–Liquiform-Labtest 87), total cholesterol (TC–Liquiform -Labtest 76) and high-density lipoprotein fraction (HDL- Liquiform-Labtest 13) were used to determinate blood lipids concentrations. Low-density lipoprotein fraction was calculated using Friedewald equation (LDL = Total Cholesterol - HDL – [Triglycerides/5]). Glycerol and FFA was also measured using an enzymatic-colorimetric methods (Free Glycerol Determination Kit - Sigma-Aldrich; Wako HR series NEFA-HR-2).
4
Measurement of Lipid Profiles
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Serum, hepatic, and muscular TAG was measured using the Sigma Triglyceride Reagent (Sigma-Aldrich, St Louis, MO, USA). Serum non-esterified fatty acid (NEFA) and glucose concentrations were measured using Wako HR Series NEFA-HR (2) and Wako Autokit Glucose (Wako Diagnostics, Mountain View, CA, USA). Serum insulin concentration was measured using the Ultra-Sensitive Mouse Insulin ELISA Kit (Crystal Chem, Chicago, IL, USA). Total lipids from the liver and hamstring muscle were extracted using isopropanol. After centrifugation, TAG content of the supernatant determined with the Sigma Triglyceride Reagent (Sigma-Aldrich, St Louis, MO, USA).
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