Native PAGE methods/recipes

[R. Jayakumar]

it is exactly same as SDS-PAGE except, do not use SDS in anything including the buffers and gel.  Moreover, you shoudl run it at 200-400V depending upon the resolution you need.  While running, You need to cool the gel plates and tank for which you need an electrophoresis aparatus with a cooling chamber through which a circulating bath circulates water at 4C.  The cold temperature is necessary to prevent the gel from heating up as well as to mainting the activity of the enzymes being seperated.  You need to do the whole thing from loading the samples to checking activity on the desired substrate quite fast.
    bye
best of luck
jayakumar

  ----- Original Message ----- 
  From: Kevin Brady 
  To: methods at hgmp.mrc.ac.uk 
  Sent: Wednesday, July 11, 2001 4:20 PM
  Subject: Native PAGE methods/recipes


  Hi,  I'm looking for a good protocol for doing Native PAGE on a protein I've purified. The theoretical pI (as calculated at Expasy ProtParam) is 7.03. Only ever done SDS-PAGE, so any help would be appreciated on gel compositions and buffers. 
  Cheers 
  Kevin 
-- 
Dr Kevin Brady
Organic Chemistry Section
School of Chemistry
University of Nottingham
Nottingham
UK
NG7 2RD
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Email : Kevin.Brady at nottingham.ac.uk or
        kevin at basil.chem.nott.ac.uk
URL   : http://www.nottingham.ac.uk/~pcxsc/thomas
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