May 15, 2019
UC Davis - IN-VIVO Glucose-stimulates Insulin Secretion Test
- Fawaz G. Haj1
- 1University of California, Davis
- Mouse Metabolic Phenotyping CentersTech. support email: info@mmpc.org
External link: https://mmpc.org/shared/document.aspx?id=86&docType=Protocol
Protocol Citation: Fawaz G. Haj 2019. UC Davis - IN-VIVO Glucose-stimulates Insulin Secretion Test. protocols.io https://dx.doi.org/10.17504/protocols.io.yptfvnn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 28, 2019
Last Modified: May 15, 2019
Protocol Integer ID: 20947
Keywords: in-vivo glucose-stimulates insulin secretion test, alterations in insulin secretion
Abstract
Summary:
An in-vivo glucose-stimulates insulin secretion test is designated to determine alterations in insulin secretion by the pancreas upon a bolus IP Glucose injection.
Materials
MATERIALS
45% Glucose solutionFisher ScientificCatalog #NC0025179(50-165-7017 replaced)
Insulin SyringesFisher ScientificCatalog #14-826-79
Saline SolutionFisher ScientificCatalog #L97753
Ultra Sensitive Mouse Insulin ELISA kitCrystal chemCatalog #90080
Heparin SodiumAbraxisCatalog #401586B
Easy Check Glucose test stripsJRS MedicalCatalog #00-101( new SKU 88982400)
Easy Check Glucose monitorJRS MedicalCatalog #Y4209 (new SKU 88972401)
Dilute the glucose stock solution (45%) with saline to 20% by adding 20ml stock to 25ml 0.9% (w/v) sterile saline.
Note:
Fisher Scientific, RRID:SCR_008452
Fast mice for 16 hours by taking away food the day before (3:00pm)
The following day, Calibrate the glucose meter according to the manufacturer’s instructions.
Deprive mice from water then measure blood glucose level using a glucometer and remove immediately approximately 50 μl of blood from the tail via a tail tip cut and transfer directly onto a sterile 0.5ml microcentrifuge tube containing 2ul Heparin.
Centrifuge at 8000rpm for 5min then transfer plasma (supernatant) to a new 0.5ml microcentrifuge tube and freeze at -80°C.
Give the mouse an intraperitoneal injection of Glucose (2g/kg) with a 27 G needle
Continue to take blood samples from the initial tail cut at 2, 5, 15 and 30 min flowing injection and repeat step3 and 4.
NOTE:
At the end of the experiment, wipe tail with 70% alcohol and allow drying. Ensure that blood loss from the tail stopped before placing the animal back to its cage.