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Tris Glycine gel running settings


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#1 Aris

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Posted 27 March 2009 - 07:11 AM

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

#2 Dr Teeth

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Posted 27 March 2009 - 07:36 AM

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.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
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#3 Aris

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Posted 27 March 2009 - 08:00 AM

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.


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???

#4 Dr Teeth

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Posted 27 March 2009 - 08:05 AM

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.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#5 Aris

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Posted 27 March 2009 - 08:23 AM

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.



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.




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