Glucagon synthetic peptide was purchased from Sigma (St. Louis, MO, USA; Cat#G2044). GLP-1 (1–37) and GLP-1(7–36 Amide) were from Bachem (Torrance, CA, USA; Cat#H-5552 and Cat#H-6795, respectively). Glucagon was reconstituted in 0.05M Acetic Acid and GLP-1 peptides were reconstituted in water at 1 mg/ml. Aliquots were stored at − 20°C until further use.
Glp 1 7 36 amide
Manufactured by Bachem
Sourced in Germany
GLP-1 7-36 amide is a synthetic peptide that corresponds to the 7-36 amino acid sequence of the endogenous glucagon-like peptide-1 (GLP-1) hormone. It is a commonly used reagent in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Glp 1 1 37, by Bachem (1 mentions)
Pyy3 36, by Bachem (1 mentions)
Ketaminol vet, by MSD (1 mentions)
Rompun vet, by Bayer (1 mentions)
Ketaminol vet, by MSD animal health (1 mentions)
Cyn 154806, by Bio-Techne (1 mentions)
Lsm 710 microscope, by Zeiss (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Plan apochromat 20x 0.8 n a objective, by Zeiss (1 mentions)
Accu chek advantage glucometer, by Roche (1 mentions)
Humulin, by Eli Lilly (1 mentions)
9 protocols using glp 1 7 36 amide
1
Glucagon and GLP-1 Peptide Preparation
2
Investigating GLP-1 Effects on Brain Metabolism
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After an overnight fasting period of 16 h, mice were icv injected with 2 μl of either saline (control) or 1 μg of GLP-1 (GLP-1 7-36 amide; #H-6795, Bachem, Switzerland) 5 min before injecting the [18F]FDG and starting the PET scans as outlined below. Icv GLP-1 or saline administration with subsequent PET imaging was conducted in a crossover fashion with a one week wash-out period between the experiments.
3
Peptide Hormone Preparation and Verification
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PYY3–36 and GLP-17–36 amide were purchased from Bachem. Following initial high-fidelity synthesis, the peptide hormones underwent purification by high-resolution, high-performance liquid chromatography. Peptide compositions and purity were verified by quantitative amino acid analysis.
Sterile 0.9% (w/v) saline was purchased from Bayer. Using an aseptic technique in a laminar flow cabinet, PYY3–36 and GLP-17–36 amide were separately dissolved in 0.9% saline, aliquoted into vials, and freeze dried. Representative PYY3–36 and GLP-17–36 amide vials were sterile after culture for 7 days (Department of Microbiology, Hammersmith Hospital, London, United Kingdom), and endotoxin levels as measured by the Limulus Amoebocyte Lysate test (Associates of Cape Cod) were within the safe range for human infusion. Further representative vials of both PYY3–36 and GLP-17–36 amide were randomly selected and sent for amino acid analysis by Alta Bioscience to calculate the actual peptide content of the vials. The bioactivity of the peptides was verified by measuring the suppression of food intake over 24 hours when injected sc into C57/BL6 mice (12 (link)), and by receptor-binding affinity assays using membranes prepared from HEK293 cells overexpressing recombinant human Y2 or GLP-1 receptor (25 (link)).
Sterile 0.9% (w/v) saline was purchased from Bayer. Using an aseptic technique in a laminar flow cabinet, PYY3–36 and GLP-17–36 amide were separately dissolved in 0.9% saline, aliquoted into vials, and freeze dried. Representative PYY3–36 and GLP-17–36 amide vials were sterile after culture for 7 days (Department of Microbiology, Hammersmith Hospital, London, United Kingdom), and endotoxin levels as measured by the Limulus Amoebocyte Lysate test (Associates of Cape Cod) were within the safe range for human infusion. Further representative vials of both PYY3–36 and GLP-17–36 amide were randomly selected and sent for amino acid analysis by Alta Bioscience to calculate the actual peptide content of the vials. The bioactivity of the peptides was verified by measuring the suppression of food intake over 24 hours when injected sc into C57/BL6 mice (12 (link)), and by receptor-binding affinity assays using membranes prepared from HEK293 cells overexpressing recombinant human Y2 or GLP-1 receptor (25 (link)).
4
Gastric Emptying and GLP-1 Infusion
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Healthy men aged 18–35 years were eligible, and those with diabetes (HbA1c >6.0% or 42.1 mmol/mol), impaired renal function or anemia, currently smoking, consuming >20 g/day alcohol, receiving medication known to affect gastrointestinal motility or glycemia, or with a history of gastric or small intestinal surgery were excluded.
Each subject attended the hospital after an overnight fast on two occasions separated by at least 4 days to be studied under regimens A and B in a randomized, double-blind fashion. Randomization was carried out by the Royal Adelaide Hospital Pharmacy using a Web-based program. Allocation concealment was maintained throughout. An intravenous catheter was inserted into each arm for drug delivery and blood sampling. Gastric emptying was measured at approximately the same time of day in each subject. During study visits, energy intake was standardized and subjects remained sitting or lying, unless toileting. GLP-1 (7–36)amide (Bachem, Germany) was infused at a rate of 0.8 pmol/kg/min, which is known to result in receptor stimulation representative of pharmacological agents (11 (link)). Placebo was 0.9% sodium chloride, and all study drugs were infused at 1 mL/min.
Each subject attended the hospital after an overnight fast on two occasions separated by at least 4 days to be studied under regimens A and B in a randomized, double-blind fashion. Randomization was carried out by the Royal Adelaide Hospital Pharmacy using a Web-based program. Allocation concealment was maintained throughout. An intravenous catheter was inserted into each arm for drug delivery and blood sampling. Gastric emptying was measured at approximately the same time of day in each subject. During study visits, energy intake was standardized and subjects remained sitting or lying, unless toileting. GLP-1 (7–36)amide (Bachem, Germany) was infused at a rate of 0.8 pmol/kg/min, which is known to result in receptor stimulation representative of pharmacological agents (11 (link)). Placebo was 0.9% sodium chloride, and all study drugs were infused at 1 mL/min.
5
Pancreas Perfusion for GLP-1R Assessment
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Pancreas perfusions were performed as previously described (46 (link)). In short, the mice were anesthetized with intraperitoneal injection of ketamine (90 mg/kg Ketaminol vet, MSD Animal Health) and xylazine (10 mg/ml, Rompun vet, Bayer Animal Health).
The stomach, kidney, and spleen were tied off. Proximally to the celiac artery, the aorta was ligated, and a catheter was inserted in the aorta thereby providing arterial perfusion with a modified Krebs-Ringer bicarbonate buffer (in mM: 118.3 NaCl, 3.0 KCL, 2.6 CaCl2*2H2O, 1.2 KH2PO4, 1.2 MgSO*2H2O, 25.0 NaHCO3, 10 glucose, 0.1% bovine serum albumin, 5% dextran) (Pharmacosmos). Effluent samples were collected through a portal vein catheter every minute. The perfusion system (UP-100 universal perfusion system, Hugo Sachs Electronic) had a constant flow of 1 mL/min, perfusion buffer was maintained at 37°C, oxygenated with 95% O2 to 5% CO2, and perfusion pressure (40-50 mmHg) was monitored throughout the experiment. GLP-1R KO mice or WT littermates (n = 8) were stimulated for 10 minutes with 0.1 nM and 1.0 nM GLP-1 7-36 amide (Bachem) at 15 and 40 minutes, respectively. At the end of the experiments, L-arginine was added as a positive control (10 mM).
Insulin concentrations in venous effluents were quantified by use of an in-house radioimmunoassay, employing ab code 2006-3 (47 , 48 (link)).
The stomach, kidney, and spleen were tied off. Proximally to the celiac artery, the aorta was ligated, and a catheter was inserted in the aorta thereby providing arterial perfusion with a modified Krebs-Ringer bicarbonate buffer (in mM: 118.3 NaCl, 3.0 KCL, 2.6 CaCl2*2H2O, 1.2 KH2PO4, 1.2 MgSO*2H2O, 25.0 NaHCO3, 10 glucose, 0.1% bovine serum albumin, 5% dextran) (Pharmacosmos). Effluent samples were collected through a portal vein catheter every minute. The perfusion system (UP-100 universal perfusion system, Hugo Sachs Electronic) had a constant flow of 1 mL/min, perfusion buffer was maintained at 37°C, oxygenated with 95% O2 to 5% CO2, and perfusion pressure (40-50 mmHg) was monitored throughout the experiment. GLP-1R KO mice or WT littermates (n = 8) were stimulated for 10 minutes with 0.1 nM and 1.0 nM GLP-1 7-36 amide (Bachem) at 15 and 40 minutes, respectively. At the end of the experiments, L-arginine was added as a positive control (10 mM).
Insulin concentrations in venous effluents were quantified by use of an in-house radioimmunoassay, employing ab code 2006-3 (47 , 48 (link)).
6
Myocardial Performance during Hyperglycemia and GLP-1
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Two separate studies were undertaken:1 ).![]()
2 ).![]()
Study 1: effects of hyperglycemia on myocardial performance during dobutamine stress
Flow chart illustrating the study design and timeline of study 1.
Study 2: effects of GLP-1 (7-36) on myocardial performance during dobutamine stress in the setting of hyperglycemia
Flow chart illustrating the study design and timeline of study 2.
7
Islet Cell Experiments with GLP-1 Analogs
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GLP‐1 (7–36) amide was from Bachem (Weil am Rhein, Germany). We used GLP‐1 at a concentration of 10 nM, in keeping with other islet cell studies (Bode et al. 1999 ; Tsuboi et al. 2003 ; de Heer et al. 2008 ; De Marinis et al. 2010 ; Shigeto et al. 2015 ; Traub et al. 2017 ). ω‐Agatoxin was purchased from the Peptide Institute (Minoh‐shi Osaka, Japan), 8‐Br‐Rp‐cAMPS (Rp‐cAMPS) from BioLog Life Science Institute (Bremen, Germany) and CYN154806 from Tocris Bioscience (Bristol, UK). Adrenaline, diazoxide, exendin (9–39), S961, and forskolin were all from Sigma‐Aldrich Company Ltd. (Gillingham, UK). When test substances were dissolved in DMSO, an equal concentration (<0.1% v/v) of the solvent was present under all control conditions.
8
Imaging Islet NAD(P)H Dynamics
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All imaging experiments were conducted in a microfluidic device at 37°C and 5% CO2. The imaging buffer consisted of (in mM) 125 NaCl, 5.7 KCl, 2.5 CaCl2•2 H2O, 1.5 MgCl2, 10 HEPES, and 0.1% bovine serum albumin (BSA; Sigma Aldrich) at pH 7.4. NAD(P)H autofluorescence was imaged with a LSM710 microscope (Carl Zeiss) using a Plan-Apochromat 20x/0.8 NA objective and a Coherent Chameleon laser tuned to 710 nm, as previously described [17] . The laser power at the sample was below 3.5 mW to prevent damage to the islet [18] (link). NAD(P)H autofluorescence was measured in intact islets as a function of glucose concentration (2–23 mM) with and without Kisspeptin-10 (KP; 1 µM; Tocris, Cat. #2570) and GLP-1 7–36 amide (20 nM; Bachem, H-6795). Addition of increasing glucose concentrations with and without the GPCR ligands occurred at 8 min intervals to allow NAD(P)H levels to plateau, after which Z-stacks were collected.
9
Insulin, GLP-1, and Intralipid Interaction
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Insulin, GLP-1, and Intralipid + heparin were infused for various times as detailed in each study protocol. Insulin (Humulin; Eli Lilly and Company) was infused systemically at 3 mU/kg/min with arterial blood glucose determined every 10 min using an Accu-Chek Advantage glucometer (Roche Diagnostics, Indianapolis, IN) and 30% dextrose (30% weight for volume) infused at a variable rate to maintain blood glucose within 10% of basal. The time course and area under the curve (AUC) of the glucose infusion rate (GIR; mg/kg/min) were calculated. GLP-1 (7–36) amide (Bachem Americas, Inc.) was infused continuously at 30 pmol/kg/min. Intralipid (6.6%) plus heparin (60 U/mL) was infused at 5 µl/min. At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature (8 (link),11 (link),23 (link),24 (link)), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses (8 (link),33 (link)).
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