Poll: Time and voltage for wet transfer to PVDF - (Apr/13/2011 )
I'm trying to use tris/glycine/20%MeOH (Towbin's) with 0.1% SDS to wet transfer onto PVDF. I think I'm blowing right through the membrane; my markers are not on the membrane or the gel after 100V for 2hrs.
If you use Towbin's buffer and a wet transfer system to PVDF (mine is bioRad mini):
What is your:
Voltage:
Time:
SDS concentration:
-polaris-
15 v
overnight.
0.1 % SDS
What size range is your protein/marker? Most proteins don't blow through PVDF.
-bob1-
polaris on Wed Apr 13 21:28:54 2011 said:
I'm trying to use tris/glycine/20%MeOH (Towbin's) with 0.1% SDS to wet transfer onto PVDF. I think I'm blowing right through the membrane; my markers are not on the membrane or the gel after 100V for 2hrs.
If you use Towbin's buffer and a wet transfer system to PVDF (mine is bioRad mini):
What is your:
Voltage:
Time:
SDS concentration:
Hola, your conditions are adequates for transfer, so have you check polarity, and activate PVDF with methanol For me these are the causes to fail. Buena suerte
-protolder-
bob1 on Wed Apr 13 23:35:01 2011 said:
15 v
overnight.
0.1 % SDS
What size range is your protein/marker? Most proteins don't blow through PVDF.
Markers go from 14k to 97k.
-polaris-
