This monoclonal antibody was raised against purified recombinant protein and is specific for the C-terminal region (residues 748-976). The antibody is not ideal for use in western blotting experiments but will work on slot blot applications. It is well suited for immunohistochemistry with paraffin embedded tissue using 1:25 to 150 dilutions, the high temperature unmasking procedure and standard ABC techniques. It will work with frozen tissue although acetone fixation is recommended. We generally recommend 60 min incubation at 25 C for the primary antibody. Specific conditions used in each lab may vary as will specific applications for this AB. For this reason, the conditions for detecting nuclear and nucleolar localization should be worked out emperically in each laboratory.
IgG Class: IgG3
Staining Pattern: Nuclear and nucleolar
Topo IIIa Backgound Information:
DNA topo III is a type IA topoisomerase that is similar to the prokaryotic (E. coli) topo I (omega protein). The reaction mechanisms for these two are similar. The prototypic topo I in animal cells is a type IB enzyme. Topo IIIa has been localized to chromosome 17 (17p11.2-12) and encodes a 976 residue, 110 kDa polypeptide. The biochemical features of this enzyme are not well understood although it is likely that it behaves similarly to the prokaryotic homologue (omega protein) in its strong preference for negatively supercoiled DNA substrates.
Purified antibody (50 ul) is supplied in 15 mM sodium azide in sterile water.
Material Safety Data:
THIS PRODUCE IS NOT LICENSED OR APPROVED FOR ADMINISTRATION TO HUMANS OR TO ANIMALS OTHER THAN EXPERIMENTAL ANIMALS. For Research Only.
The antibody is shipped on dry ice and should be stored at -20°C. We also recommend that the reagent be divided into smaller aliquots upon the first thaw. Avoid repeat freezing/thawing cycles.
Goulaouic et al. Purification and characterization of human DNA topoisomerase III. Nucleic Acids Research 27:2443-2450 (1999)
Li and Wang . Human DNA topo IIIa is essential in early embyrogenesis. PNAS 95:1010-1013 (1998)
Hanai et al. Human TOP3: A single copy gene encoding DNA topoisomerase II. PNAS 93:3653-3657 (1996)