Topoisomerase ICE Assay Kit | 고마바이오텍

HeLa cells were treated with 50uM VP-16 for 30’ and lysed in Buffer A. DNA from negative controls (no drug) and from drug treated cells was isolated and quantified on a NanoDrop. Indicated amounts of DNA were probed with TopoGEN polyclonal Top2a AB. The signals in the VP16 slots should be significantly higher than controls a shown. The signal ratio between the +DRUG:No drug control is called the “ICE DELTA” (or ICEΔ). The ICEΔ should be 2-6 fold or more for a valid result. 

HeLa cells were untreated (control) or treated with 50uM VP-16 or 50uM Camptothecin for 30’ while in exponential growth and lysed in Buffer A. DNA from negative controls (no drug) and from drug treated cells was isolated and quantified on a NanoDrop. Indicated amounts of DNA were probed with TopoGEN polyclonal Top1 Monoclonal (Cat TG2017-1). To show specificity HeLa cells were treated with an E. coli Top2 poison at three different concentrations shown (30 min at 37oC and lysed with Buffer A). Indicated amounts of DNA (0.5 to 2ug) were slot blotted.