beta gal substrate sensitivity in two-hybrid

Clare Robinson Robinson_Gordon_W.PRILVMS1 at msmail.bms.com
Mon Mar 25 11:53:41 EST 1996


Hi, the 4-MUG is definitely more sensitive than either X-gal or ONPG by 
a factor of 10-50.  The problems with 4_MUG are several-fold.  First, it 
diffuses rapidly once it has been cleaved so you have to look quickly on 
filters.  It also is brightest at high pH (8-10), so you may have to 
alter this from the normal B-gal preference (pH7).  Last, this cmpd does 
not readily enter yeast cells.  This all said, 4-MUG makes for a very 
sensitive in vitro (extract) detection of B-gal.  Look for newer 
substrates in the Molecular Probes catalog (503) 465-8300) that are 
permeable and cell-retained.  Good luck.





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