In my lab, members usually use large format gel when they perform SDS-PAGE.
Some papers reported that low current gave high resolution of the gels, so I tried about 10-20mA with constant current on large format gels for about 5 to 7 days (about 120 to 150 hours).
However, the resolution of the gels were okay to HMW, but it was bad to distinguish LMW glutenin subunit bands.
I noticed that many researchers are using mini gels. High current and voltage make gels hot, so it's better to low current and voltage in mini gels. But in my personal opinion, it's hard to distinguish the bands because the size is small.
Which size of gel is better to study low-molecular-weight glutenin? Mini gel or large gel?
this link is useful
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC34731/
regards
Hello,
can you be more specific about the molecular weight range for your Low MW glutenin proteins? A skim-reading of the above publication cited by Dr. Nima suggests that the proteins of the LMW class are 30-50kDa in size ; in which case 15% polyacrylamide (or 4-20% gradient) SDS-PAGE should have no problem resolving proteins on a typical laboratory mini or midi gel plate apparatus in 1-2 hours with good resolution.
If you are concerned about heat, you could run the apparatus in the cold room, pre-chill the buffer and sit the gel tank in an ice bath so that the buffer is readily able to heat-exchange.
Please discuss this here further, best wishes , Robin
The problem is that the bands slowly difuse the further they go. That's the disadvantage of long gels. Thus you need to find the compromise that works for you. Long gels are better for this, because you can turn them off earlier and check for the resolution.
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