Indirectly influencing cancer's out-of-control cell replication uni_wue NatureComms
C297Swas purified and the His-tag was removed as described above for PDXP protein with the following modifications: cultures were incubated for 20 h at 28 °C, protein overexpression was induced by addition of IPTG to a final concentration of 1 mM, and metal affinity chromatography was performed using a HisTrap excel nickel sepharose column .
High-throughput screening protocol: Compounds were screened at a final concentration of 10 µM in a total reaction volume of 30 µL in black, non-transparent 384-well assay plates . Measurements were conducted in the following assay buffer: Triethanolamine , NaCl , MgCl, DTT , Triton X-100 , pH 7.5.
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