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Intraperitoneal Glucose Tolerance Test Protocol
Materials
- Stock D-Glucose (dilute one gram into 10 ml of saline)
- OneTouch® Ultra Blood Glucose Monitoring System
- 0.5 cc insulin syringes
- 25G1/2 needle
- Stopwatch
- Scale
Method
- Mice are fasted for 6 hours by removal to a clean cage without food at the end of their dark (feeding) cycle. Fast time: 7am-1pm.
- Mice are weighed.
- After 6 hours of fasting, a fasting glucose level is obtained from venous blood from a small tail nick. It is measured with the OneTouch® Ultra Blood Glucose Monitoring System and test strips.
- 1mg/g body glucose is injected intraperitoneally.
- Blood glucose values are obtained at 15, 30, 60, and 120 minutes.
- About 50 µl of blood is obtained for insulin assay at 30 minutes time point.
* Tips: Keep track of when mice are injected by writing down the time that it was injected. Saving 1-2 minutes between mice if possible is good to get an accurate reading at 30 minutes and accurate insulin reading from 30’ bleed. 15 mice should be max for this test, although less would be better to get good blood draws during 30’ time point.
How to make Glucose solution:
1 gram of stock Glucose added to 10ml of Saline.
Dosage Calculation:
Stock D-glucose = 100mg/ml (weigh one gram of Glucose into 10ml of saline)
Dosage: 1mg/g glucose
1mg/g x 1ml/100 mg x 1000µl/1ml x weight of animal = amount of glucose to inject
or
= 10µl/g x body weight of animal = amount of glucose to inject |
