BioRad RotoforDIR

[fleming at guvax.acc.georgetown.edu]

In article <hAzrfQh.newittja at delphi.com>, newittja at delphi.com writes:
> "Steve A. Kay" <sak5y at FARADAY.CLAS.VIRGINIA.EDU> writes:
>  
>>Dear Netters,
>>
>>        Has anyone had any experience with the BioRad Rotofor
>>and/or Prep Cell protein purification systems ? We are
>>considering purchasing one and would appreciate any comments.
>  
>  
> I tested the Rotofor once with an E. coli lysate from cells ex-
> pressing my protein at moderate levels.  A MAJOR problem with
> this apparatus is that once the proteins near their pI, they
> precipitate.  I tried several of the suggested workarounds, but
> I never had any success.  We didn't buy the apparatus.
>  
> 	-John  <newittja at delphi.com>

This is indeed true, but we found with a number of E coli and S 
cerevisiae (mito) proteins that simply scrapping the protein off the 
nylon membranes and resuspending resulted in almost a quantitative 
recovery of activity. I miss not having the Rotofor -a WONDERFUL piece 
of equipment I thought. To me a big advantage was the ability to run 
sequential runs quickly (2 runs in 1 day was feasible with a bit of 
co-ordination), starting from a very crude 70% sat AmSO4 cut to a 
relatively pure (less than 10 bands on SDS-PAGE) fraction with minimal 
manipulation. If I had the money I would buy one again.
We trialed the Prep Cell and thought it worth the money. Unfortunately 
money didn't become availible before I moved on.
If you have the funds these two items seem well made and IMHO well 
worth the money. They would be a valuable addition to a protein 
purification setup. Rotofor-FPLC-PrepCell ... what a combination

Rob Powell

Usual disclaimers... No interest in Biorad, never worked for them etc