Peripheral plasma active GLP-1 was measured after an overnight fast and five minutes after oral gavage of 1 g/kg D-Glucose. Glucose was administered via a feeding tube to avoid stimulation of the taste buds of the lingual epithelium. Blood was collected into EDTA-coated Microvettes (SARSTEDT Inc., Newton, NC, USA) preloaded with an enzyme inhibitor cocktail containing 0.5 MEDTA (Fisher Scientific, Waltham, MA, USA), 283 μM aprotinin (Sigma-Aldrich, Burlington, MA, USA), 10,000 U/mL heparin (Sigma-Aldrich, Burlington, MA, USA), and 1.265 mM Diprotin A (Bachem, CA, USA). GLP-1 was measured using the Active GLP-1 (ver.2) Assay Kit from Meso Scale Discovery (Rockville, MD, USA), as previously described [23 (link)].
Active glp 1 ver 2 assay kit
Manufactured by Mesoscale
Sourced in United States
The Active GLP-1 (ver.2) Assay Kit is a laboratory tool designed to quantify the concentration of active glucagon-like peptide-1 (GLP-1) in biological samples. The kit provides the necessary reagents and protocols to perform this targeted analysis.
Automatically generated - may contain errors
Lab products found in correlation
Heparin, by Merck Group (1 mentions)
Diprotin a, by Bachem (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Edta coated microvette, by Sarstedt (1 mentions)
Sep pak c18 column, by Waters Corporation (1 mentions)
Dpp4 inhibitor, by Merck Group (1 mentions)
Phenylmethanesulfonyl fluoride solution, by Merck Group (1 mentions)
Aprotinin, by Roche (1 mentions)
2 protocols using active glp 1 ver 2 assay kit
1
Oral Glucose Stimulation of Active GLP-1
2
Plasma Collection and Analysis for Gut Hormones
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Blood samples for motilin analysis were collected in lithium heparin tubes, and acyl-ghrelin, GLP-1, SST and CCK samples were collected in EDTA-coated tubes all containing 500 kIU aprotinin/mL (Roche Applied Science, Penzberg, Germany). Additional DPP IV inhibitor (0.1 mM; Millipore, The Netherlands) was added for GLP-1 and phenylmethanesulfonyl fluoride solution (0.57 µM; Sigma-Aldrich, Steinheim, Germany) for acyl-ghrelin to reduce further degradation. Blood samples were centrifuged at 4°C for 10 minutes at 1400 X g. Acyl-ghrelin plasma was further acidified to a final concentration of 0.1 N HCl, extracted on a Sep-Pak C18 column (Waters Corporation, Milford, MA) and vacuum-dried. All plasma samples were directly stored at -80°C until they were used for analysis. Motilin, acyl-ghrelin and CCK levels were determined according to a radioimmunoassay as is described in earlier studies (10, 26, 27) . A commercial kit (EKE-060-03, Phoenix Pharmaceuticals, Burlingame, USA) measured SST levels, and GLP-1 levels was measured by the Active GLP-1 (ver. 2) Assay kit (Meso Scale Discovery, Maryland, USA).
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