Product Description
The topology of the DNA is a crucial issue in different biological processes like recombination, replication, transcription, and chromatin remolding. DNA topoisomerases are enzymes that control the amount of supercoiling in DNA. Without topoisomeras, DNA cannot replicate normally. The protein family of DNA topoisomerases is divided into two types (type I and II) based on their mechanism and physical properties. While the type I topoisomerases are monomeric and do not require ATP, the type II contain multi-subunits and depend on ATP for their activity. The type I enzymes include eukaryotic and bacterial topoisomerase I and III. The type II enzymes include bacterial DNA gyrase and topoisomerase IV and the eukaryotic topoisomerase II. Type I enzymes introduce transient single strand breaks in DNA, while type II enzymes produce transient double strand breaks in the DNA. Topo II plays important roles in synthesis and transcription of DNA as well as chromosomal segregation during mitosis. It is reported to be a sensitive and specific marker of late S-, G2- & M-phases in transformed and developmentally regulated normal cells Top II is also implicated in drug resistance of tumor cells. Applications: Suitable for use in Western Blot. Other applications not tested. Recommended Dilution: Western Blot: 2-10ug/ml Immunohistochemistry, Immunofluorescence: 5-10ug/ml, Formalin fixed and paraffin embedded sections (Boiled in 10mM citrate buffer, pH 6.0 for 10-20 min followed by cooling at 4C for 20 min). Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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