Insulin Tolerance Test
From Bridges Lab Protocols
Revision as of 14:19, 1 November 2010 by Davebridges (Talk | contribs) (updated insulin concentration)
Materials (Bring Downstairs
- Glucometer - AccuChek Advantage
- Glucose Test Strips - AccuChek Comfort Curve. Order these through Materiels Service by calling 936-6077 and ordering product 2535. For information see [here]
- Scale
- Beaker for weighing mice
- Syringes
- 0.07U/mL humulin in PBS (make as 7uL of 100 U/mL in 10 mL, sterile filtered). This will correspond to 0.7U/kg injections.
Timer
Protocol
- Remove food from mice for about 6h. Add do not feed tag to cages, or move cage to procedure room.
- Bring mice to procedure room. Put water bottle on cage.
- Weigh mice, mark tails as necessary and take fasting glucose measurement.
- Prepare glucose syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL).
- At 1 min intervals, inject appropriate amount of glucose into interperitoneal cavity of the mouse.
- Immobilize mouse and restrain tail with one hand
- Aim needle between the midline and the hip bone
- Insert syringe (do not inject) into cavity
- Pull up syringe and ensure that only air is aspirated. If urine or blood appears in the syringe, start over
- Eject syringe.
- At desired intervals (normally 15, 30, 45, 60, 75, 90 and 120 min), take blood glucose measurements from tail vein
- Analyse data by both % change from fasting glucose and absolute values
