Topoisomerase IIα (p170) (human) (recombinant)

Two buffers that are used to prepare a final 10X working stock of assay buffer are provided: Note that the final 10X Complete Buffer contains both A and B and should be made fresh daily! 1. 10X Incomplete Buffer A contains the following: 0.5M TRIS-HCl (pH 8.0), 1.2M KCl, 0.1M MgCl 2 , 5mM dithiothreitol) . [0.5mL provided] 2. 10X ATP Buffer B contains 50mM ATP in water [50μl provided]. These two buffers must be made fresh and mixed to make the final 10X Complete Topo II Assay Buffer (A+B) as follows. Simply add 0.1 volume of Buffer B to Buffer A. Prepare only as much of the Complete Topo II Assay Buffer (A+B) as needed for each day or set of experiments. For example: If you are performing 10 assays (20 μl/assay each) in a single day, you will need roughly 20 μl of the 10X Complete Buffer (20 μl/reaction x 10 reactions) . You should prepare 25 μl of 10X Complete Buffer (A+B) by mixing 25 μl of Incomplete Buffer A (-ATP) and 2.5 μl of ATP Buffer B. Note that this buffer should be kept on ice and used the same day, then discarded.