LAE Biotech LAE Biotech LAE Biotech


Human Topoisomerase IIa Antibody

(Catalog # AP007)


Specificity: Rabbit polyclonal antibody against Human topoisomerase IIa.


Antigen used for immunizations: Prokaryotic recombinant protein containing the amino acid residues 1095 to 1347 of the human topoisomerase IIa.


Storage Buffer and Reconstitution: The ammonium sulfate-precipitated whole serum is kept in PBS with 1% BSA and then lyophilized. Lyophilized antybody should be reconstituted with 100 ul of H2O before use.


Western blotting:

Working dilution on Western blotting 1:1000. 60 minutes primary antibody incubates at 25 oC (as shown in Fig).


Recommendations on use:


Typical working dilution on frozen sections 1:200. 60 minutes primary antibody incubates at 25 oC .

Positive controls: HeLa cells.


Storage and stability: Store unopened antibody at 4 oC or lower. Under these conditions, there is no significant loss in product performance up to one year from the purchase date. The reconstituted antibody is stable for at least two months when stored at 4 oC. For long term storage, it is recommended that aliquots of the antibody are frozen at -20 oC(frost-free freezers are not recommended). Repeated freezing and thawing must be avoided. Prepare working dilutions on the day of use.


Legal consideration: FOR RESEARCH USE ONLY.


Application: Topoisomerases are nuclear enzymes involved in a variety of cellular activities such as chromosome condensation, DNA replication, transcription, recombination and segregation at mitosis. Topoisomerase IIa is found in both prokaryotes and eucaryotes. This enzyme plays an important role in DNA replication and transcription by relieving interlocking constraints of DNA accumulation. Topoisomerase IIa is activated when two double helices cross over on another. The enzyme makes a DNA gate by breaking one double helix reversibly and let a second nearby double helix pass, then reseals the break and dissociates from the DNA. Topoisomerase protein levels increase following cell proliferation and cell cycle progression from G1 or S and G-2-M phase. The phosphorylation of the enzyme also change during the cell cycle. Phosphorylation and dephosphorylation of the Topoisomerase IIa is thought to regulate the enzyme activity in Drosphilia and Saccharomyces cerevisiae. Threonine 1342 in Human Topoisomerase IIa is phosphorylated throughout the cell cycle. Phosphorylation level of threonine 1342 is G-2M phase is as twice as much in G1 or S phase.



Chang Y-H, Hwang J, Shang H-F, et al. Characterization of human DNA Topoisomerase II as an autoantigen regognized by patients with IDDM. Diabetes. 45: 408-414 (1996).


Holden J A, Rahn M P, Jolles C J, et al.. Immunohistochemical detection of DNA topoisomerase I in formalin fixed, paraffin wax embedded normal tissues and in ovarian carcinomas. J Clin Pathol: Mol Pathol. 50: 247-253 (1997).


Li TK, Liu LF. Tumor cell death induced by topoisomerase-targeting drugs. Annu Rev Pharmacol Toxicol. 41:53-77 (2001).


McLeod H L, Douglas F, Oates M, et al.. Topoisomerase I and II activity in human breast, cervix, lung and colon cancer. Int J Cancer. 59: 607-611 (1994).


Shero J H, Bordwell B, Rothfield N F, et al.. High titers of autoantibodies to topoisomerase I (Scl-70) in sera from scleroderma patients. Science. 231: 737-740 (1986).