[Protein-analysis] Re: repack HR 10/30 column

rolando rolando.perdomo at infomed.sld.cu
Wed Jun 21 22:23:24 EST 2006


Thanks a lot for your reply. The column wasn't dried. I was
performing a cleaning method (suggested by Pharmacia using acetic acid
and NaOH and so on), but the gel was compressed very significantly. The
backpressure increased and therefore, I redueced more the flow. Anyway,
the gel compressed and large space from the column top appeared. When I
presume everything was lost with the column, it was stored horizontally
and the gel was uneven distributed through horizontal, but later on
swollen. I placed the column vertically, connect it to HPLC, adjust the
top piece above the gel and start flow through it. The column seems to
become better (apparently), but the length was reduced in 6 cm (from 30
cm) and unwanted backpressure was obtained at 1 ml/min (even when a
changed the top filter).  Looking forward to your reply.

DK wrote:
> In article <1150802575.703822.101180 at h76g2000cwa.googlegroups.com>, "rolando" <rolando.perdomo at infomed.sld.cu> wrote:
> >Hi everybody,
> >
> >I have a damaged superpose 6 HR 10/30 column from Amersham Pharmacia.
> >It is possible to repack the gel onto HR 10/30 column??? How to do
> >it???
>
> Pharmacia says you can never do as good a job they do.
> Maybe so. I think it's still possible to do decent job that gives
> acceptable resolution.
>
> What is the damage? If it's dried out, then the damage is
> mostly irreversible - agarose pores collapse upon drying. 
> 
> DK



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