After fasting the rats for 12 h, the blood glucose was collected at baseline and 15, 30, 60, 90, and 120 min after the administration of 20% glucose (1 g/kg) by an intragastric gavage for OGTT or after administration of human insulin (0.5 IU/kg) by an intraperitoneal injection for ITT. ITT was performed 2 days after OGTT to ensure full recovery. Glucose concentration was determined with glucometer (Roche Diagnostics, Germany). Serum glucagon-like peptide-1 (GLP-1) concentration was measured with multi-species GLP-1 total ELISA kit (Millipore, USA).
Multi species glp 1 total elisa kit
Manufactured by Merck Group
Sourced in United States
The Multi-Species GLP-1 Total ELISA kit is a quantitative immunoassay designed for the measurement of total glucagon-like peptide-1 (GLP-1) in various biological samples across multiple species. The kit utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.
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Lab products found in correlation
Rat insulin elisa kit, by Merck Group (2 mentions)
Glucometer, by Roche (1 mentions)
Dpp4 inhibitor, by Merck Group (1 mentions)
Human pyy total elisa kit, by Merck Group (1 mentions)
Bio plex, by Bio-Rad (1 mentions)
Supersignal west pico chemiluminescent substrate kit, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride pvdf membrane, by Merck Group (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Whatman, by Cytiva (1 mentions)
Rat leptin elisa, by Merck Group (1 mentions)
Rat mouse ghrelin total elisa, by Merck Group (1 mentions)
9 protocols using multi species glp 1 total elisa kit
1
Glucose and GLP-1 Dynamics in Rodents
2
Intestinal 3DG Absorption and GLP-1 Response
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After overnight fasting, saline (control) or 3DG at 50 mg/kg was administered to the rats by gastric gavage. Blood samples from aorta abdominals were collected before (0 min) and after (15, 30, 60, and 120 min) 3DG administration for the measurements of GLP-1 levels. Plasma GLP-1 (total) concentrations were measured using the Multi-Species GLP-1 Total ELISA kit (Millipore, MA, USA). The area under the curve (AUC) (0–120 min) for GLP-1 was calculated for each group of rats. Duodenum tissues after 1 hr administration of 3DG (50 mg/kg) were obtained for the measurement of 3DG contents by high-performance liquid chromatography, as described previously [16 (link)].
3
GLP-1 Secretion in STC-1 Cells
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STC-1 cells were seeded into 24-well plates at a density of 2 × 105 cells/well for 48 h. The cells were washed three times with phosphate-buffered saline (PBS) and then incubated with glucose-free DMEM (Gibco; Life Technologies Co., Grand Island, NY14072, USA). After 3 h, the medium was subsequently removed, and the cells were incubated with the indicated reagents for 1 h (see figures) under glucose-free (0 mM) DMEM or low-glucose (5.6 mM) DMEM conditions according to the method of Eiki et al. [24 (link)]. After the incubation, the medium was collected and centrifuged at 12 000 × g for 5 min at 4°C. The GLP-1 concentration (total) in the supernatant was measured using the Multi-Species GLP-1 Total ELISA kit (Millipore, MA, USA).
4
Appetite-associated hormone analysis protocol
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All blood samples were immediately transferred to disodium EDTA-treated tubes for analysis of appetiteassociated hormones. For the measure of PYY, GLP-1 and acylated ghrelin concentrations, test tubes were pretreated with the protease inhibitors DPP IV inhibitor (Millipore) and 4-(2-aminoethyl)benzenesulfonylfluoride hydrochloride (AEBSF, Alexis Biochemicals, Lausen, Switzerland). Blood was centrifuged at 906 g and at a temperature of 4°C for 15 min to isolate plasma. Plasma was separated and transferred to micro tubes for later analysis. Two micro tubes were pre-treated with hydrochloric acid (1 M, 100 µL per milliliter of plasma) to further protect acylated ghrelin from degradation. Plasma was stored at -70°C until hormone assays were conducted. Acylated ghrelin, total PYY and total GLP-1 were measured in duplicate using ELISA (Human Ghrelin(active) ELISA kit, Millipore; Human PYY(total) ELISA kit, Millipore; Multi Species GLP-1(total) ELISA kit, Millipore). The sensitivity of these ELISA kits were 8, 1.4 and 1.5 pg/mL, respectively, and the coefficient of variation values were 2.36, 5.26 and 3.28%, respectively.
5
Cytokine, Endotoxin, and GLP-1 Quantification Protocol
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For IL-6, endotoxin, and GLP-1 analyses, blood samples were collected at time 0 (fasting) and 1, 2, 4, and 6 h postprandial into S-Monovette (7.5-mL Z-gel) tubes and incubated for 25 min at room temperature. After centrifugation for 10 min at 1500 × g, serum was split into 1.5-mL aliquots and immediately snap-frozen in a mixture of dry ice and ethanol and stored at −80°C until analysis. IL-6 was measured with a high-sensitivity multiplex cytokine assay (Bio-Plex; Bio-Rad) by using the same kit lot for all samples to avoid lot-to-lot variability (28 (link)). The sensitivity of the kit, as provided by the manufacturer, was 0.2 pg/mL. The intra-assay CV was ≤5% and the inter-assay CV was ≤10%. Endotoxin was measured with commercially available Kinetic-Quantitative Chromogenic Limulus Amebocyte Lysate Assay (QCL-1000 LAL; Lonza). The assay was previously validated for human use with an intra-assay CV of 3.9 ± 0.5% and an inter-assay CV of 9.6 ± 0.8%, with the recovery rate for endotoxin noted at 82.0 ± 3.3% as previously detailed (29 (link)). Total GLP-1 was analyzed with the Multi Species GLP-1 Total ELISA kit (EZGLP1T-36K; Merck Millipore).
6
Hepatic FXR and TGR5 Regulation
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INT-767 was from Intercept Pharmaceuticals Inc. (New York, NY, USA). The primary antibodies against FXR, and β-actin were from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Anti-rabbit IgG HRP-linked antibody was from Cell Signaling Technologies (Danvers, MA, USA). Anti-TGR5 antibody was purchased from LifeSpan BioSciences, Inc. (Seattle, WA, USA). Pre-stained protein molecular weight marker was from Fermentas Life Sciences, (Thermo Fisher Scientific, Waltham, MA, USA). The bicinchoninic acid protein assay kit and Super-Signal West Pico chemiluminescent substrate kit were purchased from Pierce Biotechnology (Thermo Fisher Scientific). Polyvinylidene fluoride (PVDF) membranes and Whatman paper were from EMD Millipore (Billerica, MA, USA). High fat diets (HFD) (60% of calories derived from fat) and chow diets (10% of calories derived from fat) were purchased from Research Diets Inc. (New Brunswick, NJ, USA). The multi-species GLP-1 total ELISA kit and rat insulin ELISA kit were purchased from Merck Millipore (Billerica, MA, USA).
7
Ethanol-Acid Tissue Homogenization for Protein and GLP-1 Analysis
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An ethanol-acid solution was prepared by mixing absolute ethanol, water and 12 M HCl at a ratio of 74:25:1 (32) . Tissue samples were submerged in the ethanol acid solution (5 ml/g tissue) and homogenised at 25 000 rpm (Ultra-Turrax homogeniser T18, IKA) for 2 min. Homogenised samples were placed at 4°C for 24 h. After 24 h, aliquots (150 μl) of homogenate were stored separately at -30°C until protein analysis. The remaining homogenate was centrifuged at 2000 g for 20 min. The supernatant was collected and stored at -80°C until analysed. Protein and GLP-1 content were measured using Lowry's protein assay and Multi Species GLP-1 Total ELISA kit (EZGLP1T-36K; Merck Millipore), respectively. Homogenate samples were diluted 30-fold for protein measurement, whereas the supernatant was diluted 500-or 1000-fold (colon only) for GLP-1 measurement.
8
Postoperative GLP-1, PYY, and Ghrelin Levels
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Serum total GLP-1 and PYY levels after glucose gavage and fasting serum ghrelin were measured with the serum collected at 4 and 12 wk postoperatively. GLP-1 was measured with multi-species GLP-1 total ELISA kits (Millipore). PYY was measured with Rat Leptin ELISA (Millipore). Ghrelin was measured with Rat/Mouse Ghrelin (total) ELISA (Millipore).
9
Glucose-Induced Insulin and GLP-1 Measurement
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After intragastric gavage with 20% glucose (1 g/kg) at 0, 10, 30, 60, and 120 min, we collected blood specimens from the retrobulbar venous plexus. The blood supernatant was obtained after centrifugation and was stored at −80°C. We used Rat Insulin ELISA kits (Millipore, USA) and Multi-Species GLP-1 Total ELISA kits (Millipore, USA) to measure the serum insulin and total GLP-1, respectively.
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