How to do NATIVE PAGE?

[Rico Laage]

> via native PAGE (poly acrylamide gel electrophoresis).
> What do I have to think about when planning the experiment?
> Where are the differences between native PAGE and SDS-PAGE?
> 
> I heart that one has to calculate the isoelectric point.
> But how can I do this with a 300 aa-protein? (the sequence is known)
> 
> Thank you for your answer!
> 
> Thorsten Schmidt


The best protocols are the ones from Schägger et al, you can try CN PAGE
where your proteins run due to their pI (they have to have a low pI) or
you can use Blue Native PAGE where Coomassie-blue binds to proteins
giving them a charge.

Schägger et.al. "Analysis of Molecular...." -Analytical Biochemistry
217,  p220-230 (1994)

best way to get a  pI for a known sequence is the WWW-page of swissprot:
http://expasy.hcuge.ch/ch2d/pi_tool.html

have fun,

Rico

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