Topo IV Assay Buffer is supplied as a 5x Stock solution in two parts that must be mixed prior to use:
These two buffers must be mixed to prepare a final 5x complete Topo IV Assay Buffer (A+B) as follows: add 0.1 volume of Buffer B to 1.0 volume Buffer A. Prepare only as much of the complete Topo IV Assay Buffer (A+B) as needed for each day or set of experiments. Note that this buffer should be kept on ice and used the same day, then discarded.
Quality Control Analysis:
The complete Topo IV Assay Buffer (A+B) is optimized and tested for decatenation and relaxation activity of e. coli topo IV. The complete buffer should be made fresh for each experiment.
- Incomplete Buffer A contains the following: 200mM HEPES-KOH [pH8], 500mM potassium glutamate, 50mM magne- sium assetate, 50mM dithiothreitol, 250ug BSA/ml.
- ATP Buffer B contains 100mM ATP in water.
The two-part buffer is shipped on blue ice (or on dry ice if included with enzyme) and should be stored at -20°C. The complete Topo IV Assay Buffer (A+B) is stable for 24 hours at 4°C.
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