native PAGE gels

Jeffrey W. Chisholm JWChish at ac.dal.ca
Thu Mar 23 01:07:54 EST 1995


In article <ojar.melefors-2203951818210001 at 130.237.120.16>, 
ojar.melefors at postmac.cmb.ki.se says...
>
>In article <3jfgru$ddm at susscsc1.rdg.ac.uk>, abskhuri at reading.ac.uk (S.
>Khuri) wrote:
>
>Does anyone out there have any information about native PAGE gels?
>The books only seem to skim over the subject, are there any Idiot's Guides
>out there that I could have a look at?
>Thanks!!
>Sawsan Khuri
>
>I used native PAGE with subsequent staining for enzyme activity within the
>gel, e.g. for plant peroxidases (P. Jordan et al.,1990 Electrophoresis 11,
>653-655) or for human alkaline phosphatase (P. Jordan et al.,1992 Biochem.
>Int. 26, 97-104). You might wish to check the protocols. Nothing special:
>just the Laemmli-system without SDS, sample loading using sucrose, running
>buffer a Tris/glycine system. (Let me know if you need more information
>under e-mail hpeter at fm.ul.pt)

Good advice, I usually do the same except that I use Tris-Borate buffer. The major 
difficulties with native gels are choosing the optimum pH (8.3 seems to be a good place 
to start for most proteins) and running conditions.  Since native gels are usually run 
to equilibrium, you need significantly longer run times 24 hrs for some systems.

Jeffrey W. Chisholm
Lipoprotein Research Group
Dept. Of Biochemistry
Dalhousie University
Halifax, Nova Scotia, Can.

Email: JWChish at ac.dal.ca




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