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. Source: Human TOP2 beta is a 1626aa protein. Recombinant GST-fusion protein (C-temrinal ~300aa of TOP2 beta) was expressed in E. coli. GST-TOP2 beta fusion protein is ~60kD including GST. Applications: Suitable for use in Western Blot as a positive control for T8065-23. This preparation is not biologically inactive. It is not suitable for ELISA or other applications where native protein is required. Other applications not tested. Recommended Dilution: Western Blot: Heat once and load 10ul/lane and visualize with appropriate antibodies. If the product has been stored for several weeks, then it may be preferable to add 5ul of fresh 2X sample buffer per 10ul of solution prior to heating and loading on gels. Do not freeze, thaw, or heat repeatedly. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20C. Stable for 12 months at -20C. Reconstitute with sterile buffer or ddH2O. Aliquot to avoid repeated freezing and thawing. Store 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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