I'm trying to dramatically separate 150kd proteins from 100kd proteins in an acrylamide gel - more than what a 4% resolving gel would accomplish. Any ideas? Is this even possible? Thanks!
10% resolving gel will do a good job based on my experience. My running time will be usually at 1 hr and 15 mins.
8% work just fine. Try a lower power, like 60V with 20mAmp/gel. It takes a long time, but resolution is optimal. At least for me.
Depending on the buffer system and gel type you use you can get a different resolution or migration behavior of the proteins of interest. I prefer using Tris-Acetate Gels for proteins above - let's say - 75 kDa. You can get a nice separation using a 7.5 % TA-gel, but that also depends on the size/length of your gel and with that your range of separation.
I've tried 8% also and I feel like I get the same degree of separation as the 4%. I'm running basically everything off up to 100kd, but I'm still not getting the separation I would like.
For the transfer of an 8% gel I run for 1 hr and 10 min at 67 V, it works great everytime!
Hello
i agree with Mahmoud A and researchers . You can using either pure 8 or10% gels, but not gradient gels . also usually need to longer transference time , For 10% run for 1 hr and 15 mins , but 8 % run for 1 hr and 10 min
Good Luck
- Badges
- Science method