Ion-exchange chromatography

[Mr AN Qureshi]

[Posted and mailed]

In article <33CDE4E7.4E1B at bellvitge.bvg.ub.es>,
	Marta Puig <mpuig at bellvitge.bvg.ub.es> writes:
> Hi friends!
> 
> I am having problems with Haemophilus influenzae porin purifications
> with 
> DEAE columns. The protein leaves the columns just after the injection
> and 
> before the gradient starts. I guess it could be due to the column bad
> state, although I am not sure. If it is the case, is it possible to
> regenerate the column?

-- You can easily regenerate DEAE either its A-50 Sephadax or Cellulose.
With A-50 just wash the column with low salt buffer and equillibrate. With 
Cellulose you may have to wash the resin with 0.1M HCl once,
3-4 washes with deionized water then a wash with 0.1M NaOH, and lastly 5-6
times wash away the alkalie so that the pH comes down to 7-8. Then 
equillibrate the resin with desired low salt buffer and pack the column.
-------------------------------------------------------------------------------

Amer Naveed Qureshi
King's College London
Randall Institute                Phone No. : +44 (0) 171 465 5370
26-29 Drury Lane                   Fax   No. : +44 (0) 171 497 9078
London WC2B 5RL                    Email     : aqureshi at hgmp.mrc.ac.uk
UK