Avidin-biotin problems

Mark Molloy mmolloy at umich.edu
Thu Mar 30 17:29:04 EST 2000


Has anyone had difficulty with avidin-biotin interactions.

I am trying to purify a biotinylated peptide using a monomeric avidin
column.
The column is equilibrated in PBS, blocked with 2mM biotin, washed with
0.1M glycine pH2.8 then re-equilibrated with PBS. Sample is introduced
then washed with PBS pH 7.2 before elution with 5mM biotin in PBS.
Problem is the sample elutes in an early PBS wash and not seen in the
biotin elution indicating it does NOT bind to the avidin. Has anyone
experienced this and can suggest a remedy?

Thanks

-- 
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Mark Molloy, PhD                      
University of Michigan Medical School
Dept. Biological Chemistry              
Med Sci I, Room 3328
Ann Arbor, MI, 48109, USA

Tel and Fax: (734) 647 0951

http://www.proteome.med.umich.edu
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