[Protein-analysis] hydroxyapatite columns problems

Nicolas BAZEILLE via proteins%40net.bio.net (by nicolas.bazeille from curie.u-psud.fr)
Mon Feb 2 11:43:50 EST 2009


I'm trying to purify protein using hydroxypapatite system from bio-rad. The 
purification step work well, but the powder is very hard to regenerated.
I don't use any forbidden components in my buffer such EDTA or Tris.
I use different kinds of regeneration pathways (500 mM phosphate, 1M NaOH, 
6M guanidium chloride for short time) according to the supplier 
informations. However, after regeneration, the flow rate greatly decrease, 
and the powder seems to be damaged (it seems to be solubilize while 
solution containing phosphate). I don't want to use new product for each 
purification  Is there a way to preserve the stability of the material ?
Is somebody find the same problem?

thanks in advance.

Equipe XI
Génotoxicologie et cycle cellulaire - UMR 2027 CNRS/Institut Curie
Centre Universitaire
Bâtiments 110
91405 ORSAY Cedex

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