Procols and equipment for isoelectric focussing - (Dec/07/2007 )

Hi all,

I would like to try isoelectric focussing followed by western blot to observe isoforms of my favourite protein, but I can't seem to find any in-depth protocols anywhere. My protein is not membrane-associated or anything, it is ~90 kDa and has an isoelectric point of ~8.0.

Can anyone give me some advice on how to start? My lab has plenty of equipment related to SDS-PAGE gels and power packs that can go up to 400V, what else do I need?

Thanks

for ief you need a power supply that will go up to at least 2000 volts (and higher is better) as well as apparatus to run the gel.

are you going to run 2D or just ief?

are you going to pour or purchase your gels?

take a look at some catalogs or websites of companies that sell equipment and/or supplies for ief (biorad, gehealthcare(if they still do), invitrogen, etc).

There are many books on this subject.