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2 protocols using glp 1 7 37

1

Cell Lines and Reagents for Bioassays

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A transfected Chinese hamster ovary (CHO), Hela, a murine interleukin-3 dependent pro-B (Ba/F3), PC12, and rat basophil leukemia cell lines were obtained from Eurofins Scientific (Eurofins-Cerep, Le Bois I’Eveque, France). Buffers—Dulbecco’s modified Eagle medium (DMEM) buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, and Hank’s balanced salt solution (HBSS) buffer—were purchased from Invitrogen (Carlsbad, CA, USA). The reference agonists: 5′-N-ethylcarboxamidoadenosine (NECA), epinephrine bitartrate, epinephrine, DPDPE, CP 55940, linoleic acid, GLP-1(7–37, arginine vasopressin (AVP), and serotonin, and antagonists: ZM 241385, RX-821002, rauwolscine, naltriben mesylate, AM 281, exendin-3(9–39), [d(CH2)51, Tyr (Me)2]-AVP, (S)-WAY-100635, and GR55562) were obtained from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals and reagents purchased from Merck and Fluka were of the highest available grade unless otherwise stated.
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2

Calcium Signaling Pathway Optimization

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Dulbecco's Modified Eagle Medium (DMEM), HEPES, fetal calf serum (FCS), 2mercaptoethanol, penicillin-streptomycin, sodium pyruvate, Hank's balanced salt solution (HBSS), PBS, fura-2 acetoxymethyl ester (fura-2 AM) and trypsin-EDTA, were purchased from Life technologies, Sweden. Human transferrin, sodium selenite, bovine serum albumin (BSA) fraction V, nicotinamide, extracellular matrix gel, fibronectin from bovine plasma, GLP-1 (7-37), tolbutamide, D-glucose, and carbachol, were from Sigma-aldrich, Sweden.
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