I hoping for some advice regarding the use of sulfo-NHS diazirine crosslinker (Pierce) for in vitro protein interaction analysis.
Have tried labeling the protein (8.8kDa, 10 to 20uM conc) with 2 primary amines using 30-40X of crosslinker in PBS (1hour at 4deg). Have tried other concentration combinations as well, but this ratio yields some usable material. However majority of the protein crashes out of solution. Was wondering if anyone could guide me any tips and tricks in optimizing the yield of the crosslinked protein? Thanks in advance. -- Rohan Agashe
