Enzyme-linked immunosorbent assay (ELISA) kits were used to determine plasma insulin and leptin levels, specifically Mercodia Rat Insulin ELISA (Mercodia AB, Uppsala, Sweden) and Rat Leptin Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA). Circulating triglycerides (TG), non-esterified fatty acids (NEFA), and myo-inositol were measured by using the following enzymatic colorimetric kits: Serum Triglyceride Determination (Sigma Diagnostics, St. Louis, MO, USA) and NEFA-HR kit (Wako Chemicals GmbH, Neuss, Germany), according to manufacturer’s instructions, and myo-Inositol Assay Kit (Megazyme Ltd., Wicklow, Ireland), as previously adapted [27 (link)]. Glucose levels were measured in fresh blood by Accu-Check Glucometer (Roche Diagnostics, Barcelona, Spain).
Myo inositol assay kit
Manufactured by Megazyme
Sourced in Ireland
The Myo-Inositol Assay Kit is a quantitative biochemical test used to measure the concentration of myo-inositol in various sample types. It utilizes an enzymatic reaction to produce a colored product that can be measured spectrophotometrically, allowing for the determination of myo-inositol levels.
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3 protocols using myo inositol assay kit
1
Metabolic Biomarker Quantification Protocol
2
Microbial Growth and Metabolite Quantification
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Bacterial growth was monitored by measuring the optical density at 660 nm (OD 660 ) of the culture broth with a Miniphoto 518R spectrophotometer (Taitec, Saitama, Japan). Glucose concentration was determined using an F kit D-glucose (Roche Diagnostics, Basel, Switzerland) . Myo-inositol concentration was determined using a Myo-inositol assay kit (Megazyme International Ireland, Wicklow, Ireland). L-Lysine titer was determined as L-lysine HCl, according to the method described by Ohnishi et al. (2005) . Protein content was determined using a Bio-Rad protein assay kit (Bio-Rad Laboratories).
3
Quantifying Myo-Inositol in Rice Transgenics
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myo-inositol levels were quantified using myo-inositol assay kit (Megazyme, USA). TaMIPS-B over-expressing rice transgenics leaf tissue were homogenized in MQ water and centrifuged at max speed for 5 min and reaction was set as per the kit instructions. Absorbance of samples were measured at 492 nm and myo-inositol content was calculated by the formula: Myo-inositol = 0.2046 × ∆AInositol where ∆AInositol= Absorbance 2 – Absorbance1.
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