Glucose, insulin, total GIP, active GLP-1, CTX, BSAP, osteocalcin, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-β ligand (RANKL), sclerostin, and parathyroid hormone (PTH) were assayed at mins 0, 30, 60, and 120 of OGTT. Glucose, CTX, and BSAP assays were performed at Athens-Piedmont Medical Center, and insulin, GIP, GLP-1, osteocalcin, OPG, RANKL, sclerostin, and PTH were assayed at the University of Georgia College of Veterinary Medicine Cytometry Core. Serum glucose was measured via spectrophotometry using a Beckman Coulter AU5800 clinical chemistry analyzer (Beckman Coulter, Brea, CA). Serum CTX and BSAP were assayed via immunoassay using the Roche Cobas 602 (Roche Diagnostics, Basel, Switzerland) and Beckman Coulter DxI 800 (Beckman Coulter, Brea, CA), respectively. Insulin, GIP, and GLP-1 were assessed in duplicate via a magnetic bead-based multiplex platform (Millipore, HMHEMAG-34-K). Osteocalcin, OPG, sclerostin, and PTH were assessed in duplicate via a magnetic bead-based multiplex platform (Millipore, HBNMAG-51K), and RANKL was assessed using a single plex assay (Millipore, HRNKLMAG-51K).
Hmhemag 34k
Manufactured by Merck Group
Sourced in United States
HMHEMAG-34K is a lab equipment product from Merck Group. It is a magnetic particle-based separation system designed for high-throughput applications. The product facilitates the efficient separation and isolation of target molecules or cells from complex biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Au5800 clinical chemistry analyzer, by Beckman Coulter (1 mentions)
Hrnklmag 51k, by Merck Group (1 mentions)
Hbnmag 51k, by Merck Group (1 mentions)
Cobas 602, by Roche (1 mentions)
Dhwad0, by R&D Systems (1 mentions)
Nefa hr, by Fujifilm (1 mentions)
Magpix instrument, by Merck Group (1 mentions)
Dobr00, by R&D Systems (1 mentions)
Luminex magpix system, by DiaSorin (1 mentions)
Cobas c111 analyzer, by Roche (1 mentions)
6 protocols using hmhemag 34k
1
Metabolic and Bone Biomarker Dynamics
2
Evaluation of Metabolic Hormones in OGTT
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Evaluation of blood hormones was conducted after an overnight fast of 12 to 14 hours. An oral glucose tolerance test (OGTT) was performed approximately 1 week before MRI scanning. Participants ingested a 75 g glucose load, with blood sampling obtained via an arterialized hand vein at 0, 30, 60, 120, and 180 minutes. Plasma levels of Ghrelin, aGLP-1, total peptide tyrosine-tyrosine (PYY) and Leptin were measured using a human metabolic hormone magnetic bead panel (Cat. No. HMHEMAG-34K, Millipore) following the manufacturer’s instructions.
3
Measuring Metabolic Hormones in Fasts
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Blood draws were taken every 12 h of each fast (0, 12, 24, and 36 h). Samples were centrifuged for 15 min within 10 min of collection and then stored at −80 °F. Ghrelin, GLP-1, PYY, PP, leptin, insulin, and GIP were quantified using a standard 96-well multiplex human metabolic hormone magnetic bead panel according to the manufacturer’s instructions (EMD Millipore Corporation, Billerica, MA, USA, Catalog # HMHEMAG-34K).
4
Serum Biomarkers in Chilean Participants
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Venous blood samples were obtained at the clinical research facilities of the Red Salud UC (Santiago, Chile) and collected in vacuum tubes using sodium fluoride and EDTA as anticoagulants for glucose measurement and in serum separator gel tubes for insulin measurements. Glucose and insulin levels were measured in the Central Clinical Laboratory at Red Salud UC by the standard colorimetric glucose oxidase method (expressed in mg/dL) and by electrochemiluminescence immunoassay (expressed in μU/mL), respectively. Serum NEFA concentrations were determined by an enzymatic colorimetric method (NEFA-HR, Wako Chemicals, Richmond, VA). Leptin, adiponectin, and c-peptide concentrations were determined by radioimmunoassay (RIA) while sOb-R and high molecular weight (HMW) adiponectin by a commercially available ELISA (R&D Systems, codes: DOBR00 and DHWAD0, respectively). TNFα and MCP1 levels were measured by the multiplex technology in a MAGPIX instrument (Merck) using a commercial kit (HMHEMAG-34K). The mean ± SD concentrations of all of these variables are listed in Table 2 .
5
Plasma Biomarkers in Metabolic Studies
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Plasma glucose was measured on a COBAS c111 analyzer (Roche Diagnostics GmbH, Switzerland). Precision of the analyzer was verified by assaying control material provided by manufacturer (Precipath ® U/Precinorm ® U, Roche Diagnostics GmbH, Switzerland). Plasma glucagon, insulin, glucagon like peptide-1 active (GLP-1), and gastric inhibitory polypeptide (GIP) were analyzed using a commercially available Milliplex Map Kit, HMHEMAG-34K (EMD Millipore, Billerica, MA). All samples were run in duplicate on a Luminex Magpix System (Luminex, Corp. Austin, TX). The average coefficient of variation (CV) was 12.0%, 8.5%, 8.3%, and 12.2% for glucagon, insulin, GLP-1, and GIP; respectively. Inter-assay CV was 13.5%, 7.5%, 8.4% and 16.0% for glucagon, insulin, GLP-1, and GIP; respectively. Blood lactate concentration was determined via spectrophotometric assay [22 ] in triplicate. Intra-assay CV was 3.5%.
6
Quantification of Metabolic Hormone Secretion
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Hormone secretion was quantified by Luminex assay (Human Metabolic Hormone Magnetic Bead Panel, HMHEMAG-34K; EMD Millipore) according to the manufacturer’s standard protocol. After stimulation with norepinephrine or RV infection, supernatants were assayed for the amount of secreted amylin (active), C-peptide, ghrelin (active), GIP (total), GLP-1 (total), glucagon, IL6, leptin, MCP-1, PP, PYY (total), and TNF-α. Limits of detection of the assay are as follows: amylin, 11.81 pg/mL; C-peptide, 13.28 pg/mL; ghrelin, 9.40 pg/mL; GIP, 0.33 pg/mL; GLP-1, 1.11 pg/mL; glucagon, 12.73 pg/mL; IL6, 3.75 pg/mL; leptin, 54.16 pg/mL; MCP-1, 8.57 pg/mL; PP, 0.68 pg/mL; PYY, 11.18 pg/mL; and TNF-α, <0.14 pg/mL.
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