Tricks to eliminate non-specific binding of proteins to amylo se r esin, any ?

"Khadka, Deepak NICHD khadkad at mail.nih.gov
Sun Jan 20 17:17:34 EST 2002


I tried to elute the complex with the buffer containing 10 mM Maltose. Test
proteins (by the way, they were Dlx3, Msx-1, Dlxin-1 etc.) were absent in
the elute. That's what I do not want to see.

>Preblock your column ?

with what ? any suggestions ?

Deepak

> ----------
> From: 	Serial # 19781010
> Reply To: 	eikel at xs4all.nl
> Sent: 	Saturday, January 19, 2002 6:26 AM
> To: 	proteins at hgmp.mrc.ac.uk
> Subject: 	Re: Tricks to eliminate non-specific binding of proteins to
> amylose r esin, any ?
> 
> Elute your complex with maltose ?
> Preblock your column ?
> 
> grtz Tom
> 
> On 17 Jan 2002 16:32:55 -0000, khadkad at mail.nih.gov ("Khadka, Deepak
> (NICHD)") wrote:
> 
> >
> >I am performing binding assay of some test proteins with MBP-fusion
> protein.
> >When I try to purify the mixture by using amylose resin, most of the test
> >proteins gets stuck to the resin. My buffer (both for binding and
> washing)
> >is 50 mM PIPES (pH 7.0), 100 mM NaCl and 1 mM EDTA. Playing with the salt
> >concentration and adding BSA did not help. Any suggestions ?
> >
> >Deepak
> >NICHD/NIH
> >
> >---
> 
> 

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