IEF gels

Sarah Tilley sjt at newt.phys.unsw.edu.au
Wed Sep 29 02:26:31 EST 1993


-- 
Hi,

Can anyone help me? I am trying to set up an Isoelectric Focussing gel in order to study my protein. All I have is a recipe from a commercial catalogue (Hoeffer). This gives me  a recipe for a 3.5/10 native gel, and a 4/6 denaturing gel. I am worried in case my protein has a pI greater than 6 (Nobody seems to know what the exact pI is- I only have predicted values). I could do with some references and recipes for the technique. I am running my gels in a vertical system. In particular I would like to know h





ow important the blend of ampholytes is, how I can work out what the blend should be, whether sample buffers are freezable, and why I seem to get multiple bands in my markers when there should only be one! Thanks in advance.

**
Sarah Tilley
sjt at newt.phys.unsw.edu.au



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