WangLab:Pervanadate

Home        Protocol       

Preparation of pervanadate (Huyer G et al., JBC 1997)

1. Take one tube of 100mM sodium orthovanadate Na3VO4 (Must be white/colorless. If it shows yellow, means VO43- forms 12mer, cannot use.)
2. Prepare 20mM HEPES adjust PH to 7.3
3. Take 30% H2O2 dilute 10 times with HEPES buffer
4. Dilute 3% H2O2 further with HEPES buffer, 5.1ul 3% H2O2 + 44.9ul HEPES buffer
5. Add in 10ul 100mM Na3VO4, 940ul H2O. This will make up a 1mM pervanadate stock.
6. After 5 mins, add in a small amount of catalase. This will result in a burst of O2 bubbles. Open the lid of the tube to release the air pressure.
7. The pervanadate is good for several hours (<24 hours).