At the start of the inpatient metabolic chamber stay (Day 7), an intravenous catheter was inserted in the antecubital vein for blood sampling. Blood was drawn in the fasted state for ghrelin and PYY. Following breakfast, blood was sampled at 0830 and again every 30 minutes for 210 minutes, then at clock times 1300, 1500, 1700, 1800, 2000, and then again at 0200 and 0700 the following day (Day 8). The day 8 0700 blood was also analyzed for leptin. Radioimmunoassays were used to measure leptin (Millipore), PYY (Millipore), and total ghrelin (Millipore) by the CTRC Core Laboratory.
Total ghrelin
Manufactured by Merck Group
Sourced in United States
Total ghrelin is a laboratory product used for the measurement of total ghrelin levels in biological samples. Ghrelin is a hormone produced primarily by the stomach and plays a role in regulating appetite and energy balance. This product provides a tool for researchers to quantify total ghrelin concentrations, which can be useful in various scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Leptin, by Merck Group (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Interleukin 6 (il 6), by MultiSciences Biotech (1 mentions)
Amylin, by Merck Group (1 mentions)
Insulin, by Mercodia (1 mentions)
Fluostar omega, by BMG Labtech (1 mentions)
Leptin, by BioVendor (1 mentions)
5 protocols using total ghrelin
1
Hormonal Regulation during Metabolic Chamber
2
Plasma Hormones and Catecholamines Analysis
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Plasma ACTH and serum cortisol were determined by immunochemiluminescence methods (Immulite 2000 and 2500, Siemens Health Diagnostics, Deerfield, IL, USA). Total plasma ghrelin was measured with a sandwich ELISA kit measuring both intact and des-octanoyl forms with the lowest level of detection at 50 pg/mL for total ghrelin (Millipore, Billerica, MA, USA). The intra- and inter-assay CV's were 1.96% and 7.8%, respectively. Urine catecholamines and urinary free cortisol (UFC) were measured in 24 h-collections with high-performance-liquid chromatography, and liquid chromatography-tandem mass spectrometry, respectively.
3
Plasma Biomarkers after Capsaicin Feeding
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Plasma samples were obtained from the whole blood collected after 6 weeks of capsaicin feeding and stored at −80°C for further analysis. Levels of total GLP-1 (7–36 and 9–36) (Millipore), total ghrelin (Millipore), TNF-α (R&D Systems), IL-1β (R&D Systems) and IL-6 (R&D Systems) were measured by enzyme-linked immunosorbent assay according to manufacturer's instructions.
4
Measurement of Ghrelin, PYY, GLP-1, and IL-6 in Plasma
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Participants were seated, and approximately 5 mL of venous blood was drawn from the anterior cubital vein by venipuncture. The blood sample was collected in a heparin anticoagulant tube and centrifuged at 2000 g for 10 min at 4 °C. Then, the upper plasma layer was extracted and stored in a refrigerator at -80 °C. The total ghrelin (Millipore, Burlington, USA; % coefficient of variation (CV): 6.2%) and peptide YY (PYY) (Millipore, Burlington, USA; %CV: 4.8%) in the extracted sample were determined using radio-immunoassays, while glucagon-like peptide-1 (GLP-1) (Cusabio; %CV: 5.7%) and high sensitivity interleukin-6 (IL-6) (Multi science, Hangzhou, China; %CV: 6.7%) was measured using ELISA.
5
Evaluating Metabolic Biomarkers in Fasting and Postprandial States
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In study 2, under both overnight-fasting and postprandial conditions 12ml of venous blood was collected from the antecubital vein. Glucose was measured by the Accu-Chek Aviva glucose meter D r a f t 7 (Accu-Chek® Aviva, Mannheim, Germany). For further measurements, plasma was aliquoted, frozen and stored at -80 o C. Hormone measurements were carried out by enzyme-linked immunosorbent assay (ELISA) and plate reader (Fluostar Omega, BMG Labtech, Germany). ELISAs were carried out to measure amylin (Millipore, St. Charles, MO, USA) (intra assay CV: 12% ), insulin (Mercodia, Uppsala, Sweden) (intra assay CV: 7%), leptin (BioVendor Research and Diagnostic Products, BioVendor -Laboratorni medicina a.s., Czech Republic) (intra assay CV: 7% ), total ghrelin (Millipore; St. Charles, MO, USA) (intra assay CV: 4%), and PYY (Millipore Corporation, Billerica, MA, USA) (intra assay CV: 12%). The Homeostasis Model Assessment version 2 (HOMA2) (www.dtu.ox.ac.uk/homacalculator/) was used to calculate beta cell function, insulin resistance and insulin sensitivity. All samples were batch analysed and assayed in duplicate.
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