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How to stain PVDF membrane?

All conventional methods of staining may be employed with PVDF membranes with slight modifications as below.

  • Coomassie R-250:
    • Soak blott (after D.I. water rinse) in methanol
    • Stain PVDF membrane with 0.1% Coomassie R-250 in 40% MeOH for no longer than ONE MINUTE usually 15 to 20 seconds will suffice. (Staining for longer periods of time will result in high background and will interfere with extraction and cleavage)
    • Destain with 50% methanol, several changes
    • Rinse extensively with D.I. water
    • Cut out the band of interest
  • Ponceau S:
    • Stain the PVDF membrane in 0.25% Ponceau S in 1% Acetic acid for one to three minutes(until protein bands are visible)
    • Destain in D.I. water
    • Excise the band of interest
    • Rinse with D.I. water
  • Amido Black:
    • Soak blott after D.I. water rinse with methanol
    • Stain PVDF membrane with 0.2% Amido Black in 40% MeOH for 30 seconds to one minute
    • Destain in D.I. water with multiple changes until bands are clear and low background
    • Cut out band of interest