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 제품안내  면역학   Cancer Research  Topoisomerase Assay

Topoisomerase Drug Screening Kit

Topoisomerase의 작용에 대해 inhibition 활성을 갖는 potential compound를 스크리닝하는 제품입니다.

Topogen
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종류
  • E. coli Topoisomerase IV Drug Screening Kit
    • Cleavable complexes 형성을 촉진시키거나 Topoisomerase IV의 활성에 antagonize하는 2종류의 topoisomerase inhibitors 검출이 가능합니다.
    • 이미 알려져 있는 Topo II poison인 Ciprofloxacin이 control로써 제공됩니다.
    • E. coli topoisomerase IV enyzme은 별도 구매 가능합니다.
    • 구성
      • pHOT1 DNA, supercoiled
      • Marker DNA, Linear pHOT1
      • Incomplete Buffer A
      • ATP Buffer B contains 100 mM ATP
      • Ciprofloxacin control
      • 10% sodium dodecyl sulfate (SDS)
      • 10x gel loading buffer
      • Proteinase K
  • Topoisomerase I Drug Screening Kit
    • 특정 지점에서 DNA 결함을 일으키는 매개체를 안정화 시키거나 topoisomerase I의 촉매 활성을 억제시키는 lead compounds을 스크리닝 합니다.
    • Topo I enzyme은 제공되지 않습니다.
    • 구성
      • Supercoiled pHOT-1 DNA
      • Relaxed and nicked plasmid DNA markers
      • 10X Topoisomerase I assay/cleavage buffer
      • Sodium dodecyl sulfate (SDS) termination buffer (10%)
      • 10X gel loading dye
      • Control inhibitor (Camptothecin; lyophilized)
      • Proteinase K
  • Topoisomerase II Drug Screening Kit
    • 하나의 high affinity topo II cleavage site가 포함된 DNA substrate (pHOT1)를 사용하여 항균, 항바이러스, 항암 활성을 갖는 Topo II inhibitors를 스크리닝하기에 이상적입니다.
    • Cleavable complexes 형성을 촉진시키거나 Topoisomerase II 활성을 antagonize하는 2종류의 topoisomerase inhibitors 검출이 가능합니다.
    • Topoisomerase II는 별도 구매할 수 있습니다.
    • 구성
      • Supercoiled DNA [pHOT1 DNA]
      • Markers are linear pHOT1 DNA and open circular DNA
      • 10X Topoisomerase II assay buffer
      • Sodium dodecyl sulfate (10%)
      • 10X Gel loading buffer (bromophenol blue, glycerol)
      • Control inhibitor, Etoposide
      • Proteinase K
  • Topoisomerase II Drug Screening Kit (kDNA based)
    • 효소의 활성을 억제시키거나 (Catalytic Inhibitory Compounds, CICs) Cleavable complexes 형성을 촉진시키는 (Interfacial Poisons, IFPs) 2종류의 Topo II effectors 검출이 가능합니다.
    • CICs는 DNA substrate에 대한 접근을 차단하거나 ATPase의 활성 및 효소 회전율을 변경하는 등 많은 단계에서 효소에 영향을 미칠 수 있습니다.
    • IFPs는 DNA에서 resealing step 반응을 차단하는 또 다른 유형의 Inhibitor 입니다.
    • 구성
      • kDNA Substrate
      • Decatenated and Linearized kDNA Markers
      • 10X Topoisomerase II assay buffer
      • VP-16 (etoposide)
      • Sodium dodecyl sulfate (10%)
      • 10X Gel loading buffer (bromophenol blue, glycerol)
      • Proteinase K
적용
Figure 1. Use of the Topoisomerase I Drug Kit Reactions indicated above each lane were carried out using 5 units of TopoGEN type I topoisomerase. The reactions were incubated for 30 min at 37 C and stopped with SDS. After digestion with proteinase K (50 ug/ml, 60 min at 37 C), samples were loaded onto a 1% agarose gel. Under these conditions, with excess topo I and camptothecin, significant conversion of supercoiled (form I) DNA to open circular (OC DNA) was observed. It is important to recognize that cleavage assays will consume more enzyme than catalytic (relaxation type) assays. This is because each cleavage event consumes one enzyme molecule; i.e., the reaction requires stoichiometic amounts of topo I.
Figure 2. Use of the Topoisomerase II Drug Screening Kit The reactions indicated above each lane in the figure were carried out using the enzyme provided by TopoGEN and 1 ul of DNA (0.25 ug of form I pHOT-1, as provided in each kit). The reaction products were analyzed on a 1% agarose gel; the running buffer contained 0.5 ug/ml ethidium bromide. The gel was then destained in distilled water prior to photodocumentation. Formation of the cleavable complex (linear DNA species) is clearly visible. Additionally, relaxation of supercoiled DNA substrate (form I pRYG DNA) was inhibited in all reactions that contained VM26.
주문정보(Ordering Information)
CATNUM PRODUCT SIZE
TG1008-1E. coli Topo IV Drug Screening Kit (kDNA based)100 assays
TG1008-2E. coli Topo IV Drug Screening Kit (kDNA based)250 assays
TG1007-1E. Coli Topo IV Drug Screening Kit (plasmid based)100 assays
TG1007-2E. Coli Topo IV Drug Screening Kit (plasmid based)250 assays
TG2001G-1KITPurified E. coli DNA Gyrase Drug Screening Kit100 assays
TG2001G-3KITPurified E. coli DNA Gyrase Drug Screening Kit500 assays
TG2001G-5KITPurified E. coli DNA Gyrase Drug Screening Kit1000 assays
TG2001G-7KITPurified E. coli DNA Gyrase Drug Screening Kit2000 assays
TG1018-2Topo I Drug Screening Kit250 assays
TG1018-1Topo I Drug Screening Kit100 assays
TG1009-2Topo II Drug Screening Kit250 assays
TG1009-1Topo II Drug Screening Kit100 assays
TG1019-2Topo II Drug Screening Kit (kDNA Based)250 assays
TG1019-1Topo II Drug Screening Kit (kDNA Based)100 assays