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Rapid Detection and Quantitation of His-Tagged Proteins Purified by MagneHis™ Ni-Particles
By Laurie Engel, B.S., Sanchayita Kar, Ph.D., and Tonny Johnson, Ph.D.
Promega Corporation
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.
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