Attention: World-2DPAGE is no longer maintained.
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Amido Black
After electrotransfer, the PVDF membranes were stained in a solution containing Amido Black (0.5% w/v), isopropanol (25% v/v) and acetic acid (10% v/v) for 2 min. Destaining was done by several soakings in deionized water [3].
Coomassie Brilliant Blue R-250
After electrotransfer, the PVDF membranes were stained in a solution containing Coomassie Brilliant Blue R-250 (0.1% w/v) and methanol (50% v/v) for 15 min. Destaining was done in a solution containing methanol (40% v/v) and acetic acid (10% v/v) [3].
Colloidal Gold (Progold)
After electrotransfer, the PVDF membranes were incubated in PBS-Tween 0.5% for 30 minutes, washed 3 x 5 min. in PBS-Tween 0.5% and 1 min. in deionized water. They were stained in 100 ml of Problot solution overnight. Ponceau S
After electrotransfer, the PVDF membranes were stained in a solution containing Ponceau S (0.2% w/v) and TCA (3% v/v). Destaining was done by several soakings in deionized water [3].
Drying
The PVDF stained membranes were either air dried or dried on a 3 mm thick plate onto an heating plate at 37o C for 10 min [3].
Scanning
The Laser Densitometer (4000 x 5000 pixels; 12 bits/pixel) from Molecular Dynamics and the GS-700 from Bio-Rad have been used as scanning device. This scanners were linked to Sparc workstations and Macintosh computers.
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