David Ashford da5 at york.ac.uk
Wed Jun 10 11:29:00 EST 1998


You raise a very interesting point. Removal of O-glycans by
beta-elimination and recovery of the polypeptide in solution for further
testing has only rarely been reported.
I am looking to recover protein after beta-elimination to run on SDS-PAGE
and for biological testing.
I'd be interested to know of any references you know of or experiences that
you've had with this problem.
Also if you get any useful information sent directly could you post a
summary to the newsgroup.

David Ashford

Martin Dalziel wrote:
> Is the inclusion of alkaline dimethylsulfoxide the best way to minimise
> protein degredation during beta-elimination release of O-glycans in
> solution? I wish to study lectin binding to a glycoprotein after
> beta-elimination and need to minimise the loss of peptide structure
> which may result in loss of glycoprotein N-glycans. As I'm using a slot
> blotting method I understand that the solid phase Beta-elimination assay
> based on Immunobilon P membranes is not feasible due to the need for
> methanol transfer. Is there an alternative?

    Dr David A. Ashford                                                  
    Glycobiology: Research & Analytical, Department of Biology, 
    University of York, P.O. Box 373, York,  YO10 5YW  UK
    Tel: +44 (0)1904 434391           Fax: +44 (0)1904 434312   
    Email: da5 at york.ac.uk
    WWW: http://www.york.ac.uk/depts/biol/staff/daa/daa.htm  

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