.
hope this help you.
aaron hernandez
Universidad nacional autónoma de Mexico UNAM
De: "Bosch, Juergen"
Para: CCP4BB@JISCMAIL.AC.UK
Enviado: Domingo, 29 de enero, 2012 11:08:09
Asunto: Re: [ccp4bb] protein lost on membrane of centricon!!
Anothe
Another option I didn't mention is presoak your centricons in 30% glycerol over
night prior to usage.
Jürgen
On Jan 28, 2012, at 12:47 PM, Bosch, Juergen wrote:
Are you close to the theoretical isoelectric point of your protein ? Change pH
of buffer
Jürgen
..
Jürgen Bosch
J
Rashmi,
There are other membrane materials which might bind less. We have also found
that there seems to be binding sometimes to the plastic itself. So a
different manufacturer can help.
Also, always use same centricon for same protein. Sometimes you take a big
hit on a centricon's first
Are you close to the theoretical isoelectric point of your protein ? Change pH
of buffer
Jürgen
..
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W870
P.S. I haven't personally tried concentrating with PEG 20K but I suppose
it could work.
On Sat, Jan 28, 2012 at 12:41 PM, Cale Dakwar wrote:
>
> Hello Rashmi,
>
> How large are your protein monomer units? Are you expecting these units
> to have formed an oligomer? As a general rule of thumb
Hello Rashmi,
How large are your protein monomer units? Are you expecting these units to
have formed an oligomer? As a general rule of thumb, you want your protein
molecules to be no smaller than 3x the membrane MWCO. Perhaps all you need
is to try concentrating with a lower MWCO membrane, e.g.
