Removing fat from tissue preps?
Wolfgang Schechinger
Wolfgang.Schechinger at med.uni-tuebingen.de
Wed Jun 7 10:18:04 EST 2000
Anna,
in the case you shouldn't get a protocol by this NG,
I'd consider to remove the fat using chloroform (will form the
*lower* phase!), t-butanol, octanol or sth similar lipophilic,
immiscible with water. these agents will denature your proteins but
since you will boil them with SDS anyway it shouldn't matter.
In order to remove traces of solvent wich might interfere with
your SDS Page, a second extraction with up to 1 ml of MTBE, petrol
ether (not: petroleum), pentane, heptane or sth similar (avoid
hexane, it's neurotoxic) might be necessary. Maybe the higher alkanes
will work directly.
good luck!
Wolfgang
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Dr. Wolfgang Schechinger, Dept. of Pathobiochemistry
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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