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Biotin labeled oligos and transcription factor purification

Aaron Bell bellaaro at pop.pitt.edu
Mon Jan 31 15:49:22 EST 2000


I am investigating a DNA binding site which I know binds several Known
Transcription factors.  There is also one thus far unidentified binding
complex.  I have exhausted numerous antibodies of potential binding factors
to no avail.  I would like to try to immunoprecipitate the protein(s) so
that I can get some information about the factor(S) involved by subsequent
page analysis and even protein sequencing if I can isolate enough of it.  I
thought I might Biotinylate the oligo on the 5' ends and somehow (maybe UV)
crosslink the DNA protein complex then immunoprecipitate the binding
complexes with streptavadin. 
I would appreciate any input you may have regarding protein interactions
with biotinylated DNA, biotin labeling of DNA, and protein DNA crosslinking.
 thanks,
 
     Aaron W. Bell Ph.D.
     Univ. of Pittsburgh School of Medicine
     Department of Pathology
     Room S453 Biomedical Science Tower
     200 Lothrop Street
     Pittsburgh, PA 15261
     <bellaaro at pitt.edu>
     (412) 383-8787 or -7839
     (412) 648-1916 <fax>




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