Dear Didier and Opher, The difference in polarities can reach upto 60 units. I have tried concentrations between 1-5 nM DNA. Maybe I will give higher concentrations a shot.
Thanks! On Fri, Jul 21, 2017 at 4:02 PM, Didier Spittler <[email protected]> wrote: > Hello, > > What is your difference between the maximum and minimum value ? Have you > try to change the probe concentration? > > Best, > > Didier > > Le 21 juil. 2017 15:34, "Mohammad Khan" <[email protected]> a écrit : > > Dear all, > > I am trying to measure the difference in polarization upon the binding of > the DNA to my protein. I take 1-5 nM of Cy3-labelled DNA and add varying > dilutions of my protein to it (100 microM to 1 nM). I do get a decrease in > difference of polarization with decrease in protein concentration. However, > the results are difficult to reproduce and also vary greatly within > triplicates of an experiment. > > Similar observations have been observed by my colleagues with their > proteins. > > Are there any tips or precautions to keep in mind while setting up these > reactions? > > Looking forward for suggestions. > > Thank you. > > >
