VEGF release assay

Submitted by: Jessica Ye

Date: Nov 14, 2000

1. Isolate islets from WT or KO mice; wash thoroughly
2. Infect the islets with control or IRS2 adenovirus vector for 1 hour
3. wash islets thoroughly after virus infection
4. culture cells under hypoxic conditions (1% O2) or normal (20% O2) conditions for 24 or 48 hours in the presence or absence of growth factors
5. At the end of incubation, save both the culture medium and islet samples
6. measure VEGF release in the medium by ELSIA and total protein levels by protein assay
7. normalize release by total protein level.