In this report we describe methods for on-particle Coomassie® Brilliant Blue (CBB) and BODIPY® fluorescence dye binding assays to rapidly detect and quantitate His-tagged proteins. We used the CBB assay to monitor the expression of an induced His-tagged protein. The binding of CBB dye was proportional to the amount of protein bound to the MagneHis™ Ni-Particles. We were also able to bind BODIPY® fluorescent dye to His-tagged proteins associated with MagneHis™ Ni-Particles. Labeling with fluorescent dyes has the potential to be more sensitive than a CBB assay. Both of these assays should be amenable to high-throughput studies.
Promega Notes 84, 27–30.
Laurie Engel, Sanchayita Kar and Tonny Johnson