I have not seen this with mbp before, however you could try fret instead...
Artem On Apr 23, 2015 9:26 PM, "Pascal Egea" <pas...@msg.ucsf.edu> wrote: > Dear All, > > This is more a biochemical/biophysics question but since we need to > complete our structural analysis with functional or biophysical data, I > thought I would ask. We crystallized a protein as an MBP fusion and solved > its structure. Based on previous knowledge we know that the small protein > binds peptides. So we are now trying to measure binding of the protein to > peptides labeled with a fluorophore using fluorescence anisotropy. > Unfortunately the protein has to be used with its MBP still attached. > > We have a resilient problem with the MBP carrier. It has some ‘residual’ > fluorescence that interferes severely with our measurements (we are using > tetramethyl rhodamine as fluorescent reporter). > > I was curious to know if anyone else had encountered this problem and > figured out a solution. Any suggestion will be greatly appreciated. > > > Many thanks in advance. > > > Pascal Egea > > Assistant Professor of Biological Chemistry > > UCLA David Geffen School of Medicine >
