This monoclonal antibody was raised to human topo I (TG2005H-RC1) using standard methods. The antibody (IgG class) works very well for Western blotting experiments. The epitope has not been mapped. This product has not been tested for IP or IF applications; however, given the high specificity seen on Westerns, it should work well in these applications.
TG2012-4 contains 250 units of monoclonal antibody from a concentrated hybridoma supernatant. Unit definition is based on Western blotting units: one unit corresponds to a 1:1000 dilution of antibody solution sufficient to diluted working probe. The vial contains 250 ul of antibody at 1 unit/ul. For example, to prepare 10 ml of diluted antibody probe, at 10 ul of the stock antibody solution to 10 ml of suitable diluent (TBS for example). This will be sufficient to detect human topoisomerase I in 10 ug of a crude nuclear extract of typical cells (HeLa). Note that the unit concentration is very conservative and in many situations, higher dilutions of the product may work well. If the extracts are prepared from nuclei, proteolysis is less problematic. Procedures for making nuclear extracts are described here. Note that in most cases a standard whole cell extract will also work if used fresh extracts are prepared. A method for making fresh nuclear extracts is described here.
Typically you will detect a single prominent band of 100 kDa with fresh extracts. Since topo I can degrade, we recommend using nuclear extracts or freshly prepared whole cell extracts.
The antibody is stable at room temperature but should be stored at -20°C. The antibody is shipped at ambient temperature.