A rabbit polyclonal antibody was prepared against a pool of oligopeptides derived from the carboxy terminal region of human topoisomerase IIIb. The purity of the peptides was greater than 95% as determined by HPLC and mass spectral analysis. Approximately 10 mg of purified peptide were coupled (through a terminal cysteine thiol) to KLH with a heterobifunctional cross-linking agent MBS at a ratio of 1 part peptide to 1 part KLH. The antibody titer was determined by ELISA to be > 1:10000. This antibody is ideal for Western blotting applications but has not been tested for other applications. The dilutions of antibody required for specific detection should be optimized for the investigator’s particular system (cell line being used for example).
Polyclonal Antibody Unit Definition:
The antibody concentration is in Western blotting units; one unit corresponds to a 1:1000 dilution of the antibody required to make 1 ml of diluted probe for a routine Western blot. Thus, 10 units of antibody will make 10 ml of diluted probe.