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Topoisomerase Enzyme 이미지

Topoisomerase Enzyme

DNA 꼬임과 풀림에 관여하는 Topoisomerase에 대한 enzyme을 소개합니다.

주요 제품 


▒ Human Topoisomerase I (#TG2005H) 

  • Topoisomerase type I은 한개의 strand를 끊고 다시 연결합니다.
  • Human Topoisomerase I (Topo 1) 효소는 baculovirus system에서 발현된 후 정제하였습니다.
  • human tissue에서 추출한 천연 topo I에 비해 높은 순도와 특이활성을 가지며, 특히 100kDa의 Topo I 효소는 stable하며 proteolysis가 적습니다. 


Activity Assays:
Relaxation assays were carried out in a final volume of 25 µl in recombinant topo I reaction buffer (10X reaction buffer, supplied with this product is: 100 mM Tris-Cl, pH 7.9, 1.5 M NaCl, 1% BSA, 1 mM Spermidine, 50% glycerol).   Supercoiled plasmid DNA was included at 0.25 µg/reaction.  Reactions terminated with 5 µl (per 20 µl reaction volume) of stop buffer (5% sarkosyl, 0.0025% bromophenol blue, 25% glycerol). Reaction products were analyzed on a 1% native agarose gel.  Under these conditions, relaxation activity is easily detectable (typically we see 10-40 units of activity per ul).  


▒ Mutant Human Topoisomerase I (#TG2005H-Y723F)
  • 활성부위 tyrosine이 phenylalanine으로 변경된 mutant 타입의 Topo I enzyme입니다.
  • 이 enzyme은 DNA를 relax 시키지 못하며 6개의 histidine tag를 포함하고 있습니다.



Activity Assays:
Relaxation assays were carried out in a final volume of 25 μl in topo I reaction buffer (10X reaction buffer, supplied with this product is: 100 mM Tris-Cl, pH 7.9, 1.5 M NaCl, 1% BSA, 1 mM Spermidine, 50% glycerol).  Supercoiled plasmid DNA was included at 0.25 μg/reaction.  Reactions terminated with 5 μl (per 20 μl reaction volume) of stop buffer (5% sarkosyl, 0.0025% bromophenol blue, 25% glycerol).  Reaction products were analyzed on a 1% native agarose gel.  Under these conditions, relaxation activity was not detectable with up to 0.5 ug of mutant protein.


▒ Human Topoisomerase IIa (#TG2000H)
  • Topoisomerase type II는 두개의 strand를 다 끊어서 다시 연결합니다. 
  • Topoisomerase II의 경우 animal cell에는 Topo IIa와 Topo IIb의 두가지 타입으로 존재합니다. 
  • 정제된 Topo II 효소에는 Topo I 이나 Nuclease가 조금도 포함되어 있지 않습니다.
  • SDS-PAGE에서 검출된 주요 polypeptide는 170kDa 입니다. 

Topoisomerase II Assay Conditions:
Decatenation assays are carried out using kinetoplast DNA substrate in a final volume of 20-30 µl in topo II reaction buffer (50 mM Tris-Cl, pH 8.0, 120 mM KCl, 10 mM MgCl2. 0.5 mM ATP, 0.5 mM dithiothreitol) with 0.2 µg KDNA.  Reactions are terminated with 5 µl (per 20 µl reaction volume) of stop buffer (5% sarkosyl, 0.0025% bromophenol blue, 25% glycerol). Reaction products are analyzed on a 1% agarose gel containing 0.5 ug/ml ethidium bromide.  Resolution of a 2.5 kb DNA minicircle decatenated product can be easily monitored with a hand held UV light source while the gel is running.


▒ E. coli Topoisomerase IV (#TG2000EC)
  • Topo IV는 E.coli에서 parC와 parE에 의해 암호화되는 Type II topoisomerase로, decatenation과 chromosomal segregation (염색체 분리)에 필수적인 효소이며, 퀴놀론 계열의 항균제가 작용하는 효소이기도 합니다.
  • Topo IV는 KDNA를 분해하기로 하지만 supercoiled DNA를 이완시키기도 합니다. 



Assay Conditions:
Reactions contained the following (20 ul): 40 mM Tris-HCl (pH 7.5), 6 mM MgCl2, 10 mM DDT, 100 mM potassium glutamate, 40 uM ATP, 50 ug BSA/ml, 0.2 ug kDNA.  Reactions were terminated after 30 min at 37°C with SDS (1%) followed by proteinase K digestion (50 ug/ml, 15 min at 56°C).  Reaction products were run on a 1% agarose gel containing 0.5 ug/ml ethidium bromide (monitored with hand held UV light to resolve decatenation products), destained with distilled water (20 min room temperature) and photographed.


▒ E. coli DNA Gyrase (#TG2000G)
  • A,B subunit를 포함한 holoenzyme으로 제공되며, novel drug discovery를 위한 in vitro 실험이나 supercoiling plasmids 실험에 사용됩니다. 


▒ S. Aureus DNA Gyrase (#TG2000GSA)
  • 고도로 정제된 S. Aureus DNA Gyrase는 staphylococcus aureus의 두 유전자 GyrA 및 GyrB에 의해 암호화된 type II Topoisomserase로, DNA가 꼬이지 않도록 일시적으로 nick을 형성하여 DNA꼬인 정도를 조절하는 역할을 합니다.
  • 이 효소는 KDNA를 decatenating 및 supercoiling 하며, 이완된 플라스미드 DNA를 음으로 supercoiling합니다


주문정보

주문정보 - Cat No, PRODUCT, SIZE, 수량 등 항목으로 구성되어있습니다.
  Product Cat.No. Size Maker Qty Data Sheet MSDS
E. coli Topoisomerase IV TG2000EC-3 500 units TOPOGEN
E. coli Topoisomerase IV TG2000EC-1 250 units TOPOGEN
E. coli Topoisomerase IV TG2000EC-5 1000 units TOPOGEN
Human Topoisomerase I TG2005H-RC2 500 units TOPOGEN
Human Topoisomerase I TG2005H-RC1 250 units TOPOGEN
Human Topoisomerase I TG2005H-RC3 3000 units TOPOGEN
Human Topoisomerase IIa TG2000H-1 250 units TOPOGEN
Human Topoisomerase IIa TG2000H-3 1000 units TOPOGEN
Human Topoisomerase IIa TG2000H-2 500 units TOPOGEN
Human Topoisomerase IIa TG2000H-4 2000 units TOPOGEN
Mutant Human Topoisomerase I (Y723F) TG2005H-Y723F 10 ug TOPOGEN
Topoisomerase IIα C-terminal Peptide TG2011-2 50 ug TOPOGEN
human Topoisomerase II alpha (TOP2 alpha) Control/blocking peptide TOP2A11-P 100 ug ALPHA DIAGNOSTICS
Human Topoisomerase II beta-GST fusion protein for WB +ve control TOP2B21-C 100 ul ALPHA DIAGNOSTICS
Recombinant (Sf9) Human DNA Topoisomerase-I (Scl70, TOP1, >95%) RP-392 5 ug ALPHA DIAGNOSTICS
전체보기
Maker
TOPOGEN
Cat.No.
TG2000EC-3
Product
E. coli Topoisomerase IV
Size
500 units
Qty
Maker
TOPOGEN
Cat.No.
TG2000EC-1
Product
E. coli Topoisomerase IV
Size
250 units
Qty
Maker
TOPOGEN
Cat.No.
TG2000EC-5
Product
E. coli Topoisomerase IV
Size
1000 units
Qty
Maker
TOPOGEN
Cat.No.
TG2005H-RC2
Product
Human Topoisomerase I
Size
500 units
Qty
Maker
TOPOGEN
Cat.No.
TG2005H-RC1
Product
Human Topoisomerase I
Size
250 units
Qty
Maker
TOPOGEN
Cat.No.
TG2005H-RC3
Product
Human Topoisomerase I
Size
3000 units
Qty
Maker
TOPOGEN
Cat.No.
TG2000H-1
Product
Human Topoisomerase IIa
Size
250 units
Qty
Data Sheet
MSDS
Maker
TOPOGEN
Cat.No.
TG2000H-3
Product
Human Topoisomerase IIa
Size
1000 units
Qty
Data Sheet
MSDS
Maker
TOPOGEN
Cat.No.
TG2000H-2
Product
Human Topoisomerase IIa
Size
500 units
Qty
Data Sheet
MSDS
Maker
TOPOGEN
Cat.No.
TG2000H-4
Product
Human Topoisomerase IIa
Size
2000 units
Qty
Data Sheet
MSDS
Maker
TOPOGEN
Cat.No.
TG2005H-Y723F
Product
Mutant Human Topoisomerase I (Y723F)
Size
10 ug
Qty
Maker
TOPOGEN
Cat.No.
TG2011-2
Product
Topoisomerase IIα C-terminal Peptide
Size
50 ug
Qty
Maker
ALPHA DIAGNOSTICS
Cat.No.
TOP2A11-P
Product
human Topoisomerase II alpha (TOP2 alpha) Control/blocking peptide
Size
100 ug
Qty
Data Sheet
Maker
ALPHA DIAGNOSTICS
Cat.No.
TOP2B21-C
Product
Human Topoisomerase II beta-GST fusion protein for WB +ve control
Size
100 ul
Qty
Data Sheet
Maker
ALPHA DIAGNOSTICS
Cat.No.
RP-392
Product
Recombinant (Sf9) Human DNA Topoisomerase-I (Scl70, TOP1, >95%)
Size
5 ug
Qty
Data Sheet
전체보기

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