Mini centrifuge filters for protein binding studies

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Wed Nov 10 17:51:19 EST 1999


In article <80csh1$gqo$1 at tribune.usask.ca>, goldie at duke.usask.ca (Hugh
Goldie) wrote:
> I would like to use mini centrifuge filters for studies of small
> radioactive
> ligands binding to a purified protein.  At present I have a
> pressure cell,
> with eight chambers, which uses nitrogen gas pressure to force
> samples
> through 10,000 MW (Amicon TM10) filters; however, this is rather
> cumbersome
> and slow.
> Commercial filter units are available with 10,000 MW cut-offs;
> however, these
> are intended for _concentrating_ small protein samples.  They are
> constructed
> so that the solution being filtered will not all pass through the
> filter.
> (They have a "retentate volume" of 10 microlitres.)
> What I need are minicentrifuge tubes with zero retentate volume
> (i.e. all
> the solution passes through the filter and the protein is left
> behind with
> bound radioactive ligand.)  The total volume of my binding
> reactions is 100
> microlitres, so a 1.5 mL centrifuge tube would be a good size.
> Does anyone
> know where to obtain these?  I am trying to make some from various
> 1.5 mL
> and 0.5 mL tubes, but it is tricky.
> --Hugh
> --
> Hughes Goldie, Department of Microbiology and Immunology
> Saskatoon, Saskatchewan, Canada
> goldie at duke.usask.ca

Have you thought about or tried a spun-column for binding assays? It's
easy and cheap to use, e.g., a one ml G-50 or G-25 spun-column in a 1
cc syringe. And the separation time is only a couple of minutes.

Nick



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