Now I'm using 12 % SDS PAGE, I'd like to increase a gap between 85 kDa and 55 kDa protein bands to get a better result, Can anyone give me some suggestions about what percentage of SDS PAGE gel I can use to get distinct gaps between these two bands?
A 7.5% acrylamide gel should give you better separation, so that's what I'd try. If you are having problems with the bands being too broad/diffuse, then try running at a lower voltage too. Good luck!
Hello,
I used to run my 60-70 kDa samples on 9% polyacrylamide gels, and it always gave me good results.
Hope that helps.
10% gel is enough to seperate 55 to 85 KDa protein.Use acrylamide bis acrylamide ratio at 37.5:1 and run your gel at constant mA ,10 mA for stacking and 20 mA for running for a single gel.Use electrophoresis buffer by using 3 g of TRIS/lit,18 g of glycine/lit and 1g SDS/lit.It works v well,Good luck.
You can try running the conventional PAGE that are longer than the mini-protean for the same 12% gel. Even 15%gels are good to give this resolution. I case of Miniprotean run them at 150 volts for 1hr 15 min that will also help you to widen the gap between 55 and 80kDa.
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