IUBio

2D apparatus

Pierre Boule axel.siroy at free.fr
Wed Mar 26 16:54:48 EST 2003


hi guys,

I personally use both systems

my opinion is : both systems are similar, but the biorad system is cheaper
than the pharmacia which 1st dimension cells are freakly expensive

IEFcell (biorad) permits a better freezing of samples, but the Vh
(voltage.hours, quantity of V applied to the sample) range does not exceed
99999Vhrs, meanwhile IPGphor goes much far but  for biorad system voltage
increases to 10000 V (IPGphor: only 8000 V)

but, I don't agree Mr Buxbaum who said "once you have bought
> a system from a particular company, you are stuck with their gels and
> they know it.",
now we use strips from amersham (but biorad and sigma deals some strips too)
on both systems (2d dimension is performed with Xicells or dodecacell)

alright,
it's time to choose

pierre boule


"Dr Engelbert Buxbaum" <engelbert_buxbaum at hotmail.com> a écrit dans le
message de news: b5smfh$4l0$06$1 at news.t-online.com...
> L. Lintott wrote:
>
> > Hi all,
> > We are soon going to start doing some 2D work. I personally haven't done
> > any 2D work so I was looking for a bit of advice. I am considering the
> > PROTEAN IEF from BioRad. I have always been resonably happy with the
> > protein apparatuses from this company. Any one using this system and
> > happy or not happy with it?
> > Also, we want to fluorescently stain and will be imaging with the new
> > Typhoon from Amersham. Any suggestions for the best stains to use?
>
>
> Under all circumstances, get a horizontal IEF with precast gels with
> immobilised pH-gradients. Those are offered several companies, iirc also
> from BioRad. Personally, I have used Pharmacia's IGPhor system, with
> good results (no affiliation).
>
> The old method of tube gels has several disadvantages:
>
> - Samples are added at the acidic end in dilute phosphoric acid. Many
> proteins are insoluble under these conditions and do not enter the gel.
>
> - Gels are easily distorted when removing them from the tube, adding to
> variability between runs. Remember that IEF-gels need to be very low
> percentage and are correspondingly soft. Precast gels are backed with a
> plastic film, which makes handling a lot easier.
>
> - Sample size is much more limited in tubes than with the horizontal
> system, where you rehydrate the gel with your protein solution. That
> increases sensitivity.
>
> The only problem with precast gels is their price, once you have bought
> a system from a particular company, you are stuck with their gels and
> they know it.





More information about the Proteins mailing list

Send comments to us at biosci-help [At] net.bio.net