high molecular inhibitor of CYP1A1

Ralf Neunaber rneunabe at zedat.fu-berlin.de
Wed Aug 16 13:35:46 EST 1995


Dear Netters,

i´m trying to purify a Protein of not exactly detectable molecular
weight from the post microsomal supernatant of fish liver. The 
protein is highly effective in inhibiting the EROD (ethoxyresorufin-
o-deethylase)of microsomal fractions from different spezies. The
protein has a high tendency to aggregate and can only be kept 
soluble by adding high ammounts of detergents. On SDS PAGE the major
band is about 20 kD 
The problems are:
1. the protein is very instable even with added dertergents,
   so I can´t go further than the first purification step
   (maximum recovery 50%;- stable for a few hours).
2. the high ammounts of dertergents interfer with the enzymatic 
   assay.

Does anybody have experience with this or a similiar protein and can
give me tips how to stabilize and handle the protein or about
the use of special detergents.
I´m interested in trying immunoaffinity chromatography but we havn´t
got the equipment do do this. -> I would be very thankful if any
of you is interested in this problem and/or could make it possible
for me to use his equipment or give me a tip were to ask for it.

Please send your replys via E-mail, newsgroup or directly to

Ralf Neunaber
FU Berlin - Institut für Tierphysiologie -
Grunewaldstr. 34
12165 Berlin FRG
phone: +49 030-838 59 46
fax:   +49 030-838 46 40
 
many thanks for your interest.   Ralf Neunaber





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