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[Protein-analysis] Re: hydroxyapatite columns problems

Dr Engelbert Buxbaum via proteins%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Sun Feb 8 15:21:36 EST 2009


Am 02.02.2009, 12:43 Uhr, schrieb Nicolas BAZEILLE  
<nicolas.bazeille from curie.u-psud.fr>:

> Hi,
>
> 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)

Hydroxyapatite is a crystalline form of calcium phosphate. It is easier to  
use with a ceramic backbone (BioGel HTP). I am not sure why you use  
conditions like NaOH or guanidinium chloride, with a properly prepared  
sample that should not be necessary.


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