Source Paper
Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein–Coupled Receptor FFAR2
Gwen Tolhurst, Helen Heffron, Yu Shan Lam, Helen E. Parker, Abdella M. Habib et al.
Diabetes • 2011
Glucose-Stimulated GLP-1 Secretion In Vivo
Objective: Measure active GLP-1 secretion in response to glucose gavage in fasted ffar2 and ffar3 knockout mice versus wild-type controls following DPP4 inhibitor pretreatment
This is a Glucose-Stimulated GLP-1 Secretion In Vivo protocol using mouse as the model organism. The procedure involves 7 procedural steps, 2 equipment items, 4 materials. Extracted from a 2011 paper published in Diabetes.
Model and subjects
mouse • 129/SvEv • unknown • 3-4 months • similar body weight across groups
Study window
~30 minute study window | ~8 hours hands-on
Core workflow
Animal fasting • DPP4 inhibitor administration • Baseline blood collection
Primary readouts
- Active GLP-1 levels in plasma (baseline and 30 minutes post-glucose)
- Comparison of GLP-1 secretion between ffar2-/-, ffar3-/-, and wild-type mice
Key equipment and reagents
Use this page as an execution guide, then fall back to the source paper whenever you need exact exclusions, dosing details, or assay-specific caveats.
Confirm first
- Verify the animal model, intervention setup, and collection timepoints against the source paper.
- Check that every direct vendor link matches the exact specification your lab plans to run.
Use the page like this
- Work through the protocol steps in order and use the inline vendor chips only when you need to source or verify an item.
- Jump to Experimental Context for readouts, data shape, and analysis flow before planning downstream analysis.
Protocol Steps
Start here. The step list is optimized for running the experiment, with direct vendor links available inline when you need to source a cited item.
Animal fasting
Mice fasted for 4 hours prior to DPP4 inhibitor dosing
Note: Mice aged 3-4 months used
View evidence from paper
“Mice were fasted for 4 h and dosed per os with 20 mg/kg DPP4 inhibitor”
DPP4 inhibitor administration
Oral gavage administration of DPP4 inhibitor (cat. no. KR-62436) at 20 mg/kg
Note: Route: per os (oral gavage)
View evidence from paper
“dosed per os with 20 mg/kg DPP4 inhibitor (cat. no. KR-62436; Sigma)”
Baseline blood collection
Collect 150 µL blood from awake mice via tail bleed into EDTA-coated capillary tubes before glucose dosing
Note: Initial GLP-1 measurement timepoint
View evidence from paper
“For initial GLP-1 measurement, 150 µL blood was collected from awake mice via tail bleed into EDTA-coated capillary tubes (Bilbate) before glucose dosing”
Glucose gavage
Thirty minutes after DPP4 inhibitor dosing, administer glucose solution by oral gavage at 1.5 g/kg
Note: Route: oral gavage
View evidence from paper
“Thirty minutes later, mice were dosed by gavage with 1.5 g/kg glucose solution (Fisher Scientific)”
Terminal blood collection
Thirty minutes after glucose dosing, kill mice by CO2 inhalation and collect blood via cardiac puncture into aprotinin-containing tubes
Note: Euthanasia method: CO2 inhalation. Blood collected into tubes containing aprotinin (0.6 trypsin inhibiting units/mL)
View evidence from paper
“Thirty minutes after glucose dosing, mice were killed by CO 2 inhalation and blood was collected via cardiac puncture into tubes containing aprotinin (0.6 trypsin inhibiting units/mL)”
Plasma preparation
Centrifuge blood samples immediately and freeze plasma on dry ice
Note: Samples prepared for active GLP-1 assay
View evidence from paper
“Blood samples were centrifuged immediately, and plasma was frozen on dry ice before assay in duplicate for active GLP-1”
Active GLP-1 assay
Assay plasma samples in duplicate for active GLP-1 using MesoScale ELISA
Note: Samples assayed in duplicate
View evidence from paper
“plasma was frozen on dry ice before assay in duplicate for active GLP-1 (MesoScale, Gaithersburg, Maryland)”