nesyan, Vaheh
Subject: Re: [ccp4bb] mammalian cell culture on IMAC
Hi Vaheh-
Well (gulp) you could dilute your solution say 2-10X with buffer with no
salt. You could try small volumes at first and see how dilute the [salt]
must be before your protein sticks to your IEX column.
So, what is in th
This article discusses this effect in E. coli lysates. Please let us
know if you find something that works well with insect/mammalian
lysates or secreted proteins!
Enabling IMAC purification of low abundance recombinant proteins from
E. coli lysates
http://www.nature.com/nmeth/journal/v6/n7/full/n
We tried that trick, which works amazingly well in insect cells, in
mammalian media, and it fails. It will depend on the exact media, obviously.
Engin
On 10/8/09 1:50 PM, Matthew Franklin wrote:
The trick we used at Genentech (which I'm still using) was for
secreted insect cell proteins, but i
I had exactly the same problem. You should switch to Talon matrix
(Clontech). The cobalt ion is bound more tightly so stripping is
dramatically reduced. Furthermore, the Talon matrix is more specific
(then Ni Sepharose) for your His-tagged protein, so you have less
contaminants. I load two to
> -Original Message-
> From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
> Oganesyan, Vaheh
> Sent: Thursday, October 08, 2009 3:15 PM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: Re: [ccp4bb] mammalian cell culture on IMAC
>
> Dear All,
>
>
.
From: CCP4 bulletin board on behalf of Dima Klenchin
Sent: Thu 10/8/2009 3:54 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] mammalian cell culture on IMAC
>
>When mammalian cell culture is being loaded to GE HisTrap resin Ni ions
>are being stripped off
When mammalian cell culture is being loaded to GE HisTrap resin Ni ions
are being stripped off the resin, at least in my hands. Did any of you
have similar experience and if so what kind of work-around was found?
Volume is fairly large (3L) and concentration/dialysis have proven to
cause loss
Dear All,
When mammalian cell culture is being loaded to GE HisTrap resin Ni ions are
being stripped off the resin, at least in my hands. Did any of you have similar
experience and if so what kind of work-around was found?
Volume is fairly large (3L) and concentration/dialysis have proven to ca
