geoffk@usage.csd.unsw.oz.au (Geoff Kornfeld) (07/13/90)
We are presently using Cu/phenanthroline to footprint the promoter of the yeast LPD1 gene but have come across a region which does not give a good ladder in the absence of protein. As luck would have it, this region is a transcriptional activator binding site (we know that from other work). We would like to use the MPE-Fe footprinting system of Hertzberg & Dervan to gte around this (and to give us a more precise picture of what is happening at some other sites) but there are a few problems: 1) Is there a commercial distributer of MPE-Fe? I have been unable to find any here. 2) Have those out there who have used this method (& I hope that there are some) had to synthesise your own (if there is no commercial source) or were you able to get it from the authors? 3) I am willing to synthesise it if necessary, but the Australian branch of May & Baker does not have a listing for p-carboxymethidium chloride (5-methyl-6-(4-carboxylphenyl)- 3,8-diaminophenanthridinium chloride). [ the papers cite them as the source ]. Is ther an alternative source? I would appreciate it if anyone who has used this technique could help. Geoff Kornfeld School of Biochemistry UNSW