histag_experiment

Wulf Dirk Leuschner wdl at gen.mpib-tuebingen.mpg.de
Tue Nov 19 13:30:23 EST 1996


Torben Gjetting <tor at kennedy.dk> wrote:

>I am currently planning an experiment which allows me to purify
>my histagged protein (ca. 60kDa). 
>I have purchased the Ni-NTA agarose resin
>from Qiagen, and I am thinking about using the Bio-Spin columns from
>BIO-RAD.
>The reason for using spin columns is that I would like to be able to 
>do a series of purifications in parallel. If a better method that I 
>am not aware of is available, I will be most grateful for the advise.
>In addition, I would greatly apppreciate if someone can recommend an
>outline of the
>protocol for the purification. 

>Thank you in advance.
>Torben Gjetting

>-- 

>              ~~~
>              @-@ 
>    ------ooO--U--Ooo------
Hi Torben,

Qiagen sells spin-columns with a similar binding properties (they say
it shows slightly higher unspecific binding)  but based on a
silica matrix which should be easier to handle if you use a batch
procedure for purification. I am afraid these columns are pretty
expensive (Qiagen...). But you can also do the batch procedure with
the agarose resin. The only problem with this resin is that you can't
spin higher than 1000 g for 5 min, otherwise the bead material might
collapse - says Quiagen. But this works fine with me.

WDL.





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