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Tricine PAG


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#1 L u c a

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Posted 06 March 2012 - 12:53 AM

I've read that adding glycerol to tricine gel doesn't have an effect on separating proteins and it is used only to increase the density of solutions and facilitate gel casting [Tricine-SDS-PAGE protocol, Hermann Schägger]. But I doubt this. I made two variants of 10% Tricine gels that have only different glycerol concentration (10,6% & 8%). The gel with 8% glycerol showed bands with good resolution but the lowest band was about 10 kDa, human insulin marker didn't appear. The other gel showed insulin band and even a band below it (~3kDa), but the resolution of bands was much worse.
Maybe anyone can advise me how to solve the problem with resolution or give other good advises about tricine SDS PAGE?

#2 mdfenko

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Posted 06 March 2012 - 12:44 PM

you can add 6M urea to sharpen bands or run a gradient gel (we like 10-16 or 10-20%, depending on how low we need to go).
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