WangLab:Pervanadate

From OpenWetWare

Revision as of 14:33, 20 June 2007 by D. Knipmeyer (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search

Time to prepare

10 minutes

Ingredients

  • H2O
  • HEPES, 20 mM
  • Na3VO4 (in freezer)
  • Catalase (in freezer)
  • Hydrogen Peroxide, or H2O2, 30%.

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 VO4 forms 12mer, cannot use.)
  2. Prepare 20mM HEPES adjust PH to 7.3
  3. Take 30% H2O2 dilute 10 times with HEPES buffer, 5 ul 30% H2O2 + 45 ul HEPES buffer, mix gently.
  4. Dilute 3% H2O2 10 times further with HEPES buffer, 5 ul 3% H2O2 + 45 ul HEPES buffer, mix gently.
  5. Add in 10ul 100mM Na3VO4, 940ul H2O. Mix by gently reversing the vial. This will make up a 1mM pervanadate stock.
  6. After 5 minutes, use the tip of pipet to scoop a small amount of catalase, mix in to the pervanadate stock. 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).
Personal tools