Mouse monoclonal antibody prepared against an oligopeptide derived from the C-terminus of the p170 kDa human Topo IIa. The antibody was then affinity purified over a column containing the same peptide. The affinity purified antibody can be used to probe Western blots provided controls are included to clearly mark the position of the 170 kDa topo II. The reason for this is that human topo II is easily degraded or proteolyzed during routine extraction and manipulations; as a result, one may detect sub-bands. Heterogeneity in the SDS-PAGE pattern has been reported by several labs (Shelton et al., 1983; Halligan et al., 1985; Muller, et al., 1988) and recently proteolytic products of S. pombe topoisomerase II were reported (Shiozaki and Yanagida, 1991). Thus, the antibody may appear to be non-specific if there is extensive proteolysis. To counter this problem, a marker for human topoisomerase II is available (TG2011-3) which corresponds to the intact 170 kDa topoisomerase II. A second gene encoding han topoisomerase II (topo II-Beta or 180 kDa form) was reported (Chung et al., 1989; Tan et al., 1992). This antibody reacts specifically with the 170 kDa isoform of topo II.
Monoclonal Antibody to Human Topoisomerase II Alpha Unit Definition
The antibody concentration is in Western blotting units; one unit corresponds to a 1:1000 dilution of the antibody required to make a diluted probe for a routine Western blot. Thus, 10 units of antibody will make 10 ml of diluted probe (to be added directly to a single blot).