I am working the purification of protein for mass spec. I want separate some bands around 30 KDa with precast gel. I used Bolt™ 4-12% Bis-Tris Plus Gels (Invitrogen) in MOPS buffer. But it didn't seem to have a good separation at 30 KDa. Does anyone have the experience to separate the protein of this size? What gel do you recommend to use?
I used both MOPS and MES buffers for a dimeric protein of 20kDa(totally 40kDa) on the same type of gels you are using. My experience is MES has better separation for proteins around 30kDa.
Here is a separation chart for Bolt gels:
https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-gel-electrophoresis/protein-gels/bolt-bis-tris-plus-gels/bolt-bis-tris-gel-performance-data.html?open=migration#migration
I think I would choose a 12% gel with MOPS running buffer.
Try it with simple 14% Gel and normal Glycerol running buffer
or 10-20% Gradient gel
Here I found a interesting publication
http://www.pnas.org/content/110/39/15668
Maybe helps. Good luck
Tris -glycine 4-20 percent gel should separate nicely, and I got good mass spec readings from the gel slice.
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