Topoisomerase Assay Kits | 고마바이오텍

Human Topoisomerase I Assay Kit 

• Topoisomerase I은 ATP나 MgCl2의 결함 상태에서 supercoiled DNA의 꼬임을 풀어주는 Type Ib topoisomerase 입니다. 
• Supercoiled DNA가 없어지고 relaxed DNA가 관찰되는 것을 확인함으로써 Topo 1 활동을 측정합니다. 
• Kit 구성 : 
  -100 Units Human Top1 Enzyme (available with catalog number TG1015-1A only)
  -Supercoiled pHOT-1 DNA (25 µg in 100 µl TE buffer, or 0.25 µg/ul)
  -Relaxed pHOT-1 DNA marker, relaxed by topo I (50 ul at 0.05 µg/ml in gel loading buffer)
  -10X Topo I assay buffer (300 µl)
  -5X Stop buffer/gel loading dye (600 µl)
  -Detailed instruction manual
   -Sample data and documentation of all controls

Human Topoisomerase II Assay Kit

• Topoisomerase II는 p170 (alpha)와 p180 (beta) form으로 존재하며, 둘다 superhelical DNA와 kineotoplast DNA(kDNA)을 풀어주는 역할을 합니다. 
• Topo II enzyme이 kDNA network에 binding 하면서 decatenation에 의해 minicircular DNA로 풀리는 것을 측정합니다. 
  
• Kit 구성 : 
  - Human Top2a Enzyme (available with catalog no. TG1001-1A)
  - Kinetoplast DNA (kDNA)
  - Marker linear kDNA in gel loading buffer
  - Marker decatenated kDNA (topo II treated)
  - 10X Topoisomerase II assay buffer
  - 5X Stop buffer/ gel loading dye
  - Detailed instruction manual
 

Lane 1:    200ng linearized kDNA

Lane 2:    200ng decatenated kDNA

Lane 3:    No Topo II

Lane 4:    1:2 diluted Topo II

Lane 5:    1:4 diluted Topo II

Lane 6:    1:8 diluted Topo II

Lane 7:    1:16 diluted Topo II

Lane 8:    1:32 diluted Topo II

Lane 9:    1:64 diluted Topo II

DNA Intercalator/Unwinding Kit 

• 화합물의 DNA unwinding 유도 여부를 측정하기 위한 kit 입니다. 
• Kit 구성
  -Substrate DNA: pHOT1 DNA
  -Relaxed DNA Marker: 0.05 ug/ul in 1x gel loading buffer
  -10 x Topo I assay buffer
  -10x Gel Loading buffer
  -10% SDS
  -mAMSA Stock, Lyophilized (control intercalator)
  -Detailed Instruction Manual

SDS-K+ Precipitation Kit (in vitro)

• Topoisomerase I이나 II가 DNA와 반응하여 Topoisomerase/DNA 중간복합체를 형성하는 것을 확인하기 위한 kit 입니다.
• SDS와 KCl을 이용하여 Topo/DNA covalent complex를 정량하여, topo I inhibitor인지 topo II inhibitor인지, poison인지 등을 측정합니다.
• Kit 구성
  - Substrate DNA: Lambda DNA (ready for labeling)
  - 10 x Topo I assay buffer
  - 10 x Topo II assay buffer
  - 10% SDS
  - 10X TKE wash buffer
  - 2.5M KCl
  - Buffer A1 reaction buffer 

SDS-K+ Precipitation Kit (in vivo)

• Topoisomerase/DNA intermediate를 in vivo (endogeneous topoisomerase를 갖고 있는 cell, nuclei)에서 측정하기 위한 kit 입니다. 
• Kit Contents:
  - Camptothecin (10 mM)
  - Etoposide (10 mM)
  - 10X TKE wash buffer
  - 2.5M KCl
  - Cellular Lysis Solution
  - A Detailed Manual for the SDS-K+ Precipitation along with controls

Topoisomerase Assay Markers