Western Blotting: Difference between revisions
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#Transfer 4h at 75V (in cold room) or overnight at 35V (room temp). | #Transfer 4h at 75V (in cold room) or overnight at 35V (room temp). | ||
#Stain for total protein with Revert total protein stain on rocker for 5 minutes --when finished pour total protein stain back in bottle for later use! | #Stain for total protein with Revert total protein stain on rocker for 5 minutes --when finished pour total protein stain back in bottle for later use! | ||
# | #Rinse twice in revert wash solution (60ml MeOH, 13.4 ml Aceditc Acid, 126.6 ml Water) | ||
#Scan using licor for total protein, which will be used to normalize the blot | #Scan using licor for total protein, which will be used to normalize the blot | ||
#Rinse nitrocellulose in revert reversal solution for at least 5 and no more than 10 minutes until nitrocellulose appears clear again (.2g NaOH, 60ml MeOH, 140ml Water) | #Rinse nitrocellulose in revert reversal solution for at least 5 and no more than 10 minutes until nitrocellulose appears clear again (.2g NaOH, 60ml MeOH, 140ml Water) | ||