Tris Glycine gel running settings - (Mar/27/2009 )
Hi to all,
I am running currently Tris Glycine gels 6%. I havent used them so far, cause i have been running Bis Tris but for some reasons I had to change. Now i find the running settings pretty hard to adjust. I am running the gel at low Voltage i.e. 65 Volts, but it takes HOURS until the gel i run. Secondly when i bump it up it starts running in peculiar ways, not linearly, smile, sad etc. I really dont know if there are other settings or if there is another way to handle with Tris Glycine. All the protocols that i have so far seen use low Voltages for Tris Glycine.
Also as i mentioned with my former Gels (Bis Tris) i used protein from cell cultures and they were just fine. Now that i am using tissue samples, they dont run so nice. Is this a know problem between proteins from cell cultures and tissues?
Does anybody know some good running settings for Tris Glycine Gels?
thx million times
We have run Tris Glycine gels at over 100 V just fine. Are you using the correct sample buffer and Tris-Glycine buffer? We use different sample buffers and running buffers for Tris-glycine vs. Bis-Tris gels.
Dr Teeth on Mar 27 2009, 08:36 AM said:
Hi and thx for your responce, i used tris-glycine SDS running buffer and tris-glycine sample buffer (recipe that i found at invitrogen's site). Any Idea???
Have you tried running cell culture samples on the Tris-glycine gels to determine if it something in your tissue prep rather than the gels itself. Also, have you checked the expiration dates on the Tris-glycine gels? they expire much more rapidly than bis-tris.
Dr Teeth on Mar 27 2009, 09:05 AM said:
I haven't tried to run cell culture samples on tris-glycine gels. I have seen the difference between cell culture samples and tissue samples while using the bis-tris gels. The gels i used haven't expired. They are brand new and i always check before using them.