removing sarkosyl after lysis

[bipin k. dalmia]

On 25 Jun 1997 01:47:23 -0700, santbrin at CHEM.LEIDENUNIV.NL ("P.J. van
Santbrink") wrote:


>Is there an other way to get rid of the sarkosyl after lysis because I'm
>pretty sure it's the presence of the sarkosyl that's preventing the
>binding (control - empty vector pGEX - lysed without sarkosyl, binds
>good to the beads).

i don't think that's a very good control. try binding of the small
amount of soluble fusion protein lysed without sarkosyl to truly see
if sarkosyl is the one affecting binding. 

also, if sarkosyl is solubilizing 50% of your overexpressed protein,
why not use conventional chromatography (ion-exchange, size exclusion,
HIC) etc. to purify your fusion protein instead of affinity
chromatography.

bip


Bipin K. Dalmia, Ph.D.
Protein Expression and Purification
Pioneer Hi-Bred International, Inc.
Johnston, Iowa 50131

dalmiabk at phibred.com