I will be performing a Western blot for a ~350 kDa protein. I have read that tris-acetate gels are recommended for separating high molecular weight proteins. In my lab we pour our own polyacrylamide gels and use a Tris/glycine running buffer. What would be the advantage of purchasing a tris-acetate gel rather than using a low-percentage (6-8%) hand-made gel?
Suggest you a gradient precast gel like 4-12% or 4-15%. Bio-Rad TGX are working nicely in my hands with huge proteins.
Regards - Jan
Hi Margot, I'm amazed that somebody is still making their own gels!
Anyway, the advantage in purchasing precast gels, rather than casting your own gels, all comes down to cost.
This company sells 10 tris acetate 3-8%, 12 well gels for $142
http://www.lifetechnologies.com/order/catalog/product/EA03752BOX
If you factor in the labor and material cost of making these gels by hand, I'm sure that it will be a lot more than $14.2 per gel.
I might also add that commercial precast gels are made in bulk and are QC'd to perform as expected. Hand-made gels might, or might not work.
If the handmade gels don't work, you will spend $$$ and time figuring out what is wrong.
Hi Margot
No problem with casting your own gels - in my opinion all students should try it sometime...
I am not sure this will answer your question but I do know that the pH of Tris/glycine gels can rise to >9 during electrophoresis which can lead to deamination and alkylation. Given the longer running time to separate very large proteins, this may become an issue. As I recall it the pH of Tris acetate gels are closer to pH 7.
Hope this helped
Jesper
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