Re: [ccp4bb] Off-topic, protein in dye-front (ion front?) on native-PAGE

Hi All, Sorry to bring this old topic up again. I planned to run tricine gels but I found a possible error in table 2 (4% stacking gel formula) in Hermann Schägger protocol (Nature Protocols volume 1, pages 16–22 (2006), the author wrote 3ml 3X gel buffer in a total of 12 ml solution, it should

Re: [ccp4bb] Off-topic, protein in dye-front (ion front?) on native-PAGE

Try Tricine gel. It is particularly suited for low molecular wt proteins and it will give you very good resolution. Even you can run it overnight at 30 V (16-18 hours). On Thu, Jan 19, 2017 at 9:33 AM, Walt wrote: > Hi, > > I have a small protein (~9 kDa) with acidic pI (~4).

Re: [ccp4bb] Off-topic, protein in dye-front (ion front?) on native-PAGE

A) change dye or use pure glycerol to load gel B) change gel pH or add various substances to it (easy to do with the buffer) Artem On Jan 19, 2017 6:34 AM, "Walt" wrote: > Hi, > > I have a small protein (~9 kDa) with acidic pI (~4). > When I run 18% native-PAGE, it appears

[ccp4bb] Off-topic, protein in dye-front (ion front?) on native-PAGE

Hi, I have a small protein (~9 kDa) with acidic pI (~4). When I run 18% native-PAGE, it appears my protein is in the dye front. How can I fix this problem? Changing the pH of separating gel might help? How about gradient native-PAGE? Thank you! Walt