Blood was drawn after an overnight fast. Plasma glucose was quantified (YSI-2300, Yellow Springs Instruments) and subjects classified as normal (NG; FG<100mg/dL) or impaired glycemia (IG; FG≥100mg/dL) based upon the American Diabetes Association cut-point for impaired fasting glucose. Twelve subjects who met the cut-point for impaired glycemia had a prior diabetes diagnosis and all diabetic subjects were on diabetic medication. We also quantified additional metabolic biomarkers (Insulin (Genway), amylin (Millipore) and non-esterified fatty acids (NEFA, Wako Diagnositcs)) in plasma using enzyme linked immunosorbent assay. Body mass was assessed using a digital scale accurate to 0.1kg (Seca Platform Scale, model 707). HOMA-IR was calculated from the product of glucose and Insulin divided by 405.
Amylin
Manufactured by Merck Group
Sourced in United States
Amylin is a laboratory equipment product manufactured by Merck Group. It is designed to measure and analyze samples in a laboratory setting. The core function of Amylin is to provide accurate and reliable data for research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (1 mentions)
Ysi 2300, by Yellow Springs Instruments (1 mentions)
Ghrelin, by Merck Group (1 mentions)
Rmhmag 84k, by Merck Group (1 mentions)
Luminex 200, by DiaSorin (1 mentions)
Srage, by R&D Systems (1 mentions)
Total ghrelin, by Merck Group (1 mentions)
Insulin, by Mercodia (1 mentions)
Fluostar omega, by BMG Labtech (1 mentions)
Leptin, by BioVendor (1 mentions)
Chrysin, by Merck Group (1 mentions)
Congo red, by Merck Group (1 mentions)
Thioflavin t, by Merck Group (1 mentions)
6 protocols using amylin
1
Metabolic Biomarkers in Glycemia
2
Multiplex Profiling of Rat Metabolic Hormones
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In vivo rat study: Plasma concentrations of peptide hormones were determined using a customized xMAP based multiplex-assay (Milliplex map rat metabolic hormone magnetic bead panel – metabolism multiplex assay, cat. no. RMHMAG-84 K, Millipore), for the following selected analytes: Amylin (Active), C-Peptide, Ghrelin (Active), GIP (Total), GLP-1 (total), Glucagon, IL-6, Insulin, Leptin, MCP-1, PP, PYY (Total), TNF-α. The assay is reported to have no significant cross-reactivity with other hormone tested. After the analytes were bound to the antibody-coupled beads, a biotinylated detection antibody was added, followed by incubation with streptavidin-phycoerythrin conjugate. Subsequently, the beads were subjected to a flow cytometry-based detection method using a Luminex laser-based analyzer (catalog no. Luminex 200, Luminex, Austin, TX 78727, USA). The manufacturer's instructions were closely followed. Reported intra- and inter-assay variation was 1–8% and 7–29%, respectively. The median fluorescent intensities from the eight calibrators were used to interpolate concentrations in plasma samples using 5-parameter logistic regression.
3
Serum Biomarkers Analysis Protocol
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Participants reported to the clinic following an overnight fast, and blood was collected via venipuncture into serum vacutainer tubes containing clot activator. Blood was processed for serum, and glucose was measured using a YSI 2300 Stat Plus Glucose Lactate Analyzer. All remaining serum was stored at −80°C until further analysis. Insulin (Genway), amylin (Millipore), S100B (Millipore), sRAGE (R&D), and esRAGE (As One International) were quantified using ELISA per the manufacturer's protocol. Serum cRAGE was computed by subtracting esRAGE from the total plasma sRAGE pool as previously described [33 (link)], and the cRAGE : esRAGE ratio was subsequently calculated [29 (link)].
4
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.
5
Amyloid Inhibitor Screening Protocol
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Chrysin (5,7-Dihydroxyflavone), Amylin, Congo red, Thioflavin T was purchased from Sigma-Aldrich, Inc. St Louis, MO, USA.
6
Culturing Human Fibroblasts and Derivatives
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Cell culture-media/conditions, drugs Human fibroblasts (HFTE, primary human lung fibroblasts Clonetics CC2512 immortalized with hTERT), WI38, and 293T cells and derivatives were cultured at 37 C and 5% CO 2 in DMEM with 10% FBS. All cells were cultured at 37 C and 5% CO 2 . 4-OHT (LH6278; Sigma-Aldrich) was dissolved in EtOH, purified amylin (A5972; Sigma-Aldrich) was dissolved in DMSO.
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