Gut Protein extracts (+DNA)

Mr SMNN Faruque s.faruque at umds.ac.uk
Wed Feb 21 14:20:29 EST 1996


I am studying DNA with transcription factor binding sites using EMSA'a 
("gel-shifts").  
My crude protein preps from whole embryos work well, but when I employed 
this method on adult gut (duodenum, jejunum, ileum and posterior colon) 
I not only am unable to see binding even with an Sp1 concensus binding 
site probe, but when added to embryo protein in a binding reaction it 
inhibited binding (or degraded the binding proteins?).
I suspected the presence of active proteases in the gut extracts, but 
boiling of the sample for 10 minutes at 100degC prior to addition to the 
binding mixture did not stop this effect.

If anyone has anyideas or experience with gut protein extracts, please 
mail me.

Nadeem



      Nadeem Faruque             Department of Craniofacial Development
      Tel 0171-955 4782                           Floor 28, Guy's Tower
      Fax 0171-955 2704	                              UMDS, SE1 9RT, UK
                <http://www.umds.ac.uk/elsewhere/cfd/cfd.html>



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