problem with elution from hydrophobic column

Dan Diaz bl275 at cleveland.Freenet.Edu
Fri Mar 12 22:58:15 EST 1993


In a previous article, blatch at uctvax.uct.ac.za () says:

>PROBLEM WITH ELUTION CONDITIONS FOR A HYDROPHOBIC COLUMN:
>
>We are trying to purify a protein using a hydrophobic material called FRACTOGEL
>TSK BUTYL-650 (M) produced by Merck. The matrix consists of butyl residues
>linked by an ether link to a polyvinyl support matrix. Our protein sample will
>stick to the matrix at approx 0.8 M ammonium sulphate but under no
>circumstances will any proteins elute from the matrix even if we use distilled
>water. Note our protein sample will not stick to the matrix in the absence of
>salt. So we are dealing with what seems to be an irreversible binding process.
>What are we doing wrong?  Must we just change to a completely different
>hydrophobic moiety? Any suggestions will be greatly appreciated. Please send
>suggestions to me or Neil at micro.uct.ac.za
>
>
>
>

Suggestions:
1. Add ethylene glycol to you eluent
2. Lower the temperature (weakens Hphobic interactions)
3. Use a less hydrophobic resin, like phenyl
-- 
Dizzy Dan        Department of Molecular Genetics and Curried Sauces
     Albert Einstein College of Medicine, Da Bronx, New York



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