Abstract We are requesting an Administrative Supplement for Equipment Purchases to obtain an AKTA Pure 25M Fast Protein Liquid Chromatography (FPLC) System to replace our failing AKTA Purifier FPLC system installed in 2007. The requested instrument meets all of our specific needs and will maintain compatibility with our existing chromatography media and methods. The AKTA Pure 25M is capable of detecting multiple wavelengths, which is essential when working with the fluorescent dyes needed for single molecule experiments. The proposed system is capable of the full range of chromatography methods in particular the low pressure columns we use for small scale purifications. The ability to generate recombinant protein samples of high purity and then separate them from fluorescent dyes forms the basis of all experiments under the parent award. This award was funded for 4 years starting 09/20/2023.