Plasma insulin levels were determined using ELISA colorimetric insulin assay kit (Crystal Chem). For iron measurement, plasma samples were collected from heparinized blood and used for measurement of total circulating iron (Fe2+ and Fe3+) using the QuantiChrom Iron Assay Kit (BioAssay Systems). Plasma hepcidin levels were determined using Hepcidin-Murine CompeteTM ELISA (Intrinsic Life Sciences). For GLP-1, whole blood samples were collected in microtubes treated with 1:10μl anti-proteolytic cocktail (EDTA/Aprotinin/Heparin) to prevent enzymatic GLP-1 degradation. Plasma total GLP-1 levels were determined using the Meso Scale Discovery Platform according to the manufacturer’s instructions (Meso Scale Discovery). All assays were performed according to the manufacturer’s instructions.
Elisa colorimetric insulin assay kit
Manufactured by Crystal Chem
The ELISA colorimetric insulin assay kit is a laboratory equipment used to quantify insulin levels in biological samples. It employs the enzyme-linked immunosorbent assay (ELISA) technique, which utilizes specific antibodies and color-changing enzymes to detect and measure the concentration of insulin.
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3 protocols using elisa colorimetric insulin assay kit
1
Plasma Biomarker Quantification Protocol
2
Glucose Tolerance and Gastric Emptying
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During glucose tolerance tests, all mice were fasted for 4–5 hr prior to oral administration of dextrose (2 g/kg body weight). Tail vein blood glucose levels were measured using Accu-Chek glucometers (Accu-Chek Aviva Plus, Roche Diagnostics) or Biosen C-line glucose analyzer (EKF diagnostic) at 0, 15, 30, 45, 60, 90, and 120 min post glucose administration. Gastric emptying rate was assessed by an oral gavage of glucose (2 g/kg) with acetaminophen (100 mg/kg) in 4–5 hr fasted mice. Blood was collected from the tail vein at baseline and 10 min after gavage in EDTA-coated microtubes. Plasma acetaminophen levels were used to assess the rate of gastric emptying and were measured using spectrophotometry assay (Sekisui Diagnostics). Plasma insulin levels were determined using ELISA colorimetric insulin assay kit (Crystal Chem).
3
Plasma Biomarker Quantification Protocol
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Plasma insulin levels were determined using ELISA colorimetric insulin assay kit (Crystal Chem). For iron measurement, plasma samples were collected from heparinized blood and used for measurement of total circulating iron (Fe2+ and Fe3+) using the QuantiChrom Iron Assay Kit (BioAssay Systems). Plasma hepcidin levels were determined using Hepcidin-Murine CompeteTM ELISA (Intrinsic Life Sciences). For GLP-1, whole blood samples were collected in microtubes treated with 1:10μl anti-proteolytic cocktail (EDTA/Aprotinin/Heparin) to prevent enzymatic GLP-1 degradation. Plasma total GLP-1 levels were determined using the Meso Scale Discovery Platform according to the manufacturer’s instructions (Meso Scale Discovery). All assays were performed according to the manufacturer’s instructions.
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