A polyclonal antibody was prepared against a 16 residue oligopeptide derived from the carboxy terminal region of human topoisomerase II. The purity of the peptide was greater than 95% as determined by HPLC and mass spectral analysis. The antibody titer was determined by ELISA to be > 1:104. Immunoaffinity gel chromatography and antibody purification steps were carried out to obtain a homogeneously purified IgG.
This antibody is ideal for either Western blotting applications or immunoflourecence (IF) studies on the cytological localization of topoisomerase II. The dilutions of antibody required for specific detection should be optimized for the investigator’s particular system (cell line being used for example); however, we supply the antibody reagent along with a detailed protocol for immunofluorescence using a typical human cell line (optimized for HeLa cells). A second reagent, the 16 -mer oligopeptide ( TG2011-2) is also available. This allows the investigator to perform proper controls for absorbing the antibody to demonstrate specificity in IF experiments. The IF protocol has been optimized for tumor cells that possess relatively high levels of topoisomerase II. In addition, we offer a Western blot marker set for topo II (TG2011-3) to unambiguously locate the position of the 170 kDa topo II