Protein ultrafiltration query

Jeffrey Yuziuk jayuziuk at rs5.tcs.tulane.edu
Fri Nov 15 12:29:38 EST 1996


Are you using cellulose based membranes? You may have a cellulase present.
Do standard proteins of similar size pass through the membranes before or
after are used for your enzyme?

Jeff

Haralt Leiter (hleiter at edv2.boku.ac.at) wrote:
: Sorry, I earlier forgot a subject line:
: 
: Hi everybody !
: 
: I´m a poor PhD Student (my supervisor hasn´t paid me) purifying a
: fucosyltransferase with an estimated (SDS-PAGE) molecular weight of at
: least 50kDa. During ultrafiltration the entire enzyme-activity passes
: through the membrane (cutoff 30000). Has anybody observed a similar
: effect or can offer a reasonable explanation ?
: I use a 25mM Tris/HCl pH7,3
:         0,1% Triton X-100
:         10% Glycerol
:         0,02% NaN3    buffer and the applied protein solution contains
: several
: hundreds of micograms.
: Thanks for any answers in advance
: 
: Haralt
: 
: BTW, I have already tried membranes of different companies



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