IEF unit recommendations.

Deanne Bell dbell at fresno.ars.usda.gov
Fri Jun 4 11:59:11 EST 2004


Hi Sean:
My lab-buddy doing 2-D has used the multiphor II in the past, but his
current favorite is the Bio-Rad Protean IEF system.  

Why more than you asked for but - I personnally think the really crucial
element is the IMAGING:

1. HIGHLY recommend Sypro Ruby for: MUCH BETTER reproducibility; Ruby
has better linearity for quantitation; Ruby sensitivity exceeds the mass
spec sensitivity.
It's costly, but you can re-use the stain at least one time without
losing too much sensitivity.  And I don't use the recommended volume
(recommended 700 ml for a large gel...I use 400 mL).  

2. Another friend compared imagers - the Typhoon from Amersham, the
FMBIO from MiraiBio (formerly Hitachi), the ProExpress from Perkin Elmer
(Genomic Solutions), Bio-Rad's FX Pro, and Bio-Rad's FluorS MultiImager.
She says "The Typhoon absolutely blew the other imagers out of the
water.  It gave the best images and definitely the most sensitive.
MiraiBio's was a close second, but had a problem with the outer edges of
the image bleaching out.  Bio-Rad, I wouldn't even consider.  They were
SIGNIFICANTLY less sensitive.  Where we detected about 2500 spots with
the Typhoon, I think there were only about 800 detected with Bio-Rad's
system. The nice thing about the Typhoon is it can image all sorts of
fluorescent dyes for protein, RNA and DNA, but it can also do
chemilluminescence and phosphor imaging.

3. a huge evaluation of all 2DE image analysis software on the market
found The two best were Progenesis from Non-Linear Dynamics, and Protein
Mine from Scimagix.  
PDQuest and Melanie are much cheaper, but not very good.  DeCyder from
Amersham analyzes the multiplexed images of CyDyes.  It's based on
Progenesis and it's fantastic.  You can even look at rotating 3D graphs
of each individual spot.  It's also got all sorts of bells and whistles
as far as statistical tools.  

TTFN
Dee Bell
-----Original Message-----
From: owner-methods at hgmp.mrc.ac.uk [mailto:owner-methods at hgmp.mrc.ac.uk]
On Behalf Of Sean Patterson
Sent: Friday, June 04, 2004 7:39 AM
To: methods at hgmp.mrc.ac.uk
Subject: IEF unit recommendations.

Hi listers.

	Do people have comparative experience/recommendations for the 
purchase of an IEF unit for 2D separations? We're going to buy a 
system, and basically we're looking at the Amersham Pharmacia 
Multiphor or Ettan IPGphor systems, the Bio-Rad Protean IEF system, 
and the Invitrogen Zoom IPGRunner system.

	The uses would be for general 2D protein characterization, 
but eventually some proteomics work as well.

Sean
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