topogen代理

topogen代理-上海金畔

TopoGEN是一家研究诊断公司,为从事DNA拓扑异构酶和DNA修复的研究人员提供创新的产品和服务。从事基于机制的抗癌、抗病毒和抗菌药物发现的科学家会发现其产品在研发中特别有用。他们的药物筛选试剂盒可用于明确鉴定拓扑异构酶靶向分子,用于癌症化疗和抗菌应用。TopoGEN还销售检测和定量原核(DNA Gyrase,拓扑异构酶IV)和真核拓扑异构酶(拓扑异构酶I,IIa,IIb,III)的检测试剂盒。TopoGEN还提供全系列的 DNA 底物和 DNA 切割靶标、拓扑异构酶抑制剂、拓扑异构酶抗体和试剂盒补充试剂。DNA修复的相关系列产品也是他们产品的关键部分。

TopoGEN生产和销售研究高质量试剂,并了解出版物和专li所需的证据标准。TopoGEN的技术支持人员致力于为您(研究人员)提供完成研究目标所需的培训和敏锐度。

topogen常卖产品:

货号 品名 规格 品牌
DR3000 HRHeLa-Sce HR based Kit in HeLa Requiring I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR3000 HRHeLa-Sce2 HR based Kit in HeLa Requiring I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR3000 HRHeLa-Sce3 HR based Kit in HeLa Requiring I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR3000 HRHeLa-CPR HR based Kit in HeLa Requiring CRISPR Transfection (50ug DNA) 1Kit topogen
DR3000 HRHeLa-CPR2 HR based Kit in HeLa Requiring CRISPR Transfection (100ug DNA) 1Kit topogen
DR3000 HRHeLa-CPR3 HR based Kit in HeLa Requiring CRISPR Transfection (250ug DNA) 1Kit topogen
DR3000 HRCN1.4-Sce HR based Kit in Mouse Neuronal Cells Requiring I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR3000 HRCN1.4-Sce2 HR based Kit in Mouse Neuronal Cells Requiring I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR3000 HRCN1.4-Sce3 HR based Kit in Mouse Neuronal Cells Requiring I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR3000 HRSH-Sce HR in human Neuroblastoma Cells req. I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR3000 HRSH-Sce2 HR in human Neuroblastoma Cells req. I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR3000 HRSH-Sce3 HR in human Neuroblastoma Cells req. I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR3000 HR3T3-Sce HR based Kit in MOUSE 3T3 Cells  Requiring I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR3000 HR3T3-Sce2 HR based Kit in MOUSE 3T3 Cells  Requiring I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR3000 HR3T3-Sce3 HR based Kit in MOUSE 3T3 Cells  Requiring I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR3000 Custom-Sce Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.)  Includes HRHeLa cell line control (50ug DNA) 1Kit topogen
DR3000 Custom-Sce2 Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.)  Includes HRHeLa cell line control (100ug DNA) 1Kit topogen
DR3000 Custom-Sce3 Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.)  Includes HRHeLa cell line control (250ug DNA) 1Kit topogen
DR3000A Custom-Sce Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.) No HRHeLa cell line control (50ug DNA) 1Kit topogen
DR3000A Custom-Sce2 Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.) No HRHeLa cell line control (100ug DNA) 1Kit topogen
DR3000A Custom-Sce3 Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.) No HRHeLa cell line control (250ug DNA) 1Kit topogen
DR5001 HNHela-Sce NHEJ Based HeLa Requires I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR5001 HNHela-Sce2 NHEJ Based HeLa Requires I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR5001 HNHela-Sce3 NHEJ Based HeLa Requires I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR5000 HNSH-Sce NHEJ Based in Human Neuroblastoma Line requires I-Sce1 transfection (50ug DNA) 1Kit topogen
DR5000 HNSH-Sce2 NHEJ Based in Human Neuroblastoma Line requires I-Sce1 transfection (100ug DNA) 1Kit topogen
DR5000 HNSH-Sce3 NHEJ Based in Human Neuroblastoma Line requires I-Sce1 transfection (250ug DNA) 1Kit topogen
DR5000 HNCN1.4-Sce NHEJ Based in Mouse Neuronal CN1.4 cells requires I-Sce1 transfection (50ug DNA) 1Kit topogen
DR5000 HNCN1.4-Sce2 NHEJ Based in Mouse Neuronal CN1.4 cells requires I-Sce1 transfection (100ug DNA) 1Kit topogen
DR5000 HNCN1.4-Sce3 NHEJ Based in Mouse Neuronal CN1.4 cells requires I-Sce1 transfection (250ug DNA) 1Kit topogen
DR5000 HN3T3-Sce NHEJ Based Mouse 3T3 cells Requires I-Sce1 Transfection (50ug DNA) 1Kit topogen
DR5000 HN3T3-Sce2 NHEJ Based Mouse 3T3 cells Requires I-Sce1 Transfection (100ug DNA) 1Kit topogen
DR5000 HN3T3-Sce3 NHEJ Based Mouse 3T3 cells Requires I-Sce1 Transfection (250ug DNA) 1Kit topogen
DR5000 Custom-Sce Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  Includes iHNHeLa cell line control (50ug DNA) 1Kit topogen
DR5000 Custom-Sce2 Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  Includes iHNHeLa cell line control (100ug DNA) 1Kit topogen
DR5000 Custom-Sce3 Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  Includes iHNHeLa cell line control (250ug DNA) 1Kit topogen
DR5000A Custom-Sce Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  NO iHNHeLa cell line control (50ug DNA) 1Kit topogen
DR5000A Custom-Sce2 Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  NO iHNHeLa cell line control (100ug DNA) 1Kit topogen
DR5000A Custom-Sce3 Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  NO iHNHeLa cell line control (250ug DNA) 1Kit topogen
DR3050-1 HR Pooled Cell analysis:  50ug DR-HRGFP, 50ug pSce1, 25ug e-GFP 1Kit topogen
DR3050-2 HeLa Cell RepHR Pooled Cell analysis:  250ug DR-HRGFP, 250ug pSce1, 25ug e-GFP 1Kit topogen
DR3050-3 HR Pooled Cell analysis:  500ug DR-HRGFP, 500ug pSce1, 25ug e-GFP 1Kit topogen
DR3050-4 HR Pooled Cell analysis:  1000ug DR-HRGFP, 1000ug pSce1, 25ug e-GFP 1Kit topogen
DRT5050-1 Detailed protocols with 50ug of cNHEJ plasmid, 50ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRT5050-2 Detailed protocols with 250ug of cNHEJ plasmid, 250ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRT5050-3 Detailed protocols with 500ug of cNHEJ plasmid, 500ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRT5050-4 Detailed protocols with 1000ug of cNHEJ plasmid, 1000ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRA5050-1 Detailed protocols with 50ug of Alt-NHEJ plasmid, 50ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRA5050-2 Detailed protocols with 250ug of Alt-NHEJ plasmid, 250ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRA5050-3 Detailed protocols with 500ug of Alt-NHEJ plasmid, 500ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
DRA5050-4 Detailed protocols with 1000ug of Alt-NHEJ plasmid, 1000ug I-Sce1 plasmid, 50ug eGFP transfection control plasmid 1Kit topogen
TG1050-1 Topoisomerase Type IIa Nuclear Extraction/Assay Kit 100assays topogen
TG1001-1 Topoisomerase II Assay Kit (20ug kDNA) 100assays topogen














TopoGEN书2020年版来啦~~

 

 

美国TopoGEN公司成立于1991年,是一家诊断公司,由一批在拓扑异构酶领域工作和在基础、临床、应用及相关方面研究经验丰富的研究者创建,为以药物开发为基础的拓扑异构酶和机制研发人员提供创新产品和服务。药物筛选试剂盒可以分析拓扑异构酶活性药物,用于癌症化疗和抗菌方面。TopoGEN公司还能提供原核和真核化验拓扑异构酶 (I型,II,IV和旋转酶)检测试剂盒,体外体内诊断均可应用。TopoGEN公司还提供DNA载体、DNA裂解目标、拓扑异构酶抑制剂、拓扑异构酶抗体和相关试剂盒等完整产品线。

上海金畔TopoGEN书2020年版来啦~~

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TopoGEN书

美国TopoGEN公司成立于1991年,是一家诊断公司,由一批在拓扑异构酶领域工作和在基础、临床、应用及相关方面研究经验丰富的研究者创建,为以药物开发为基础的拓扑异构酶和机制研发人员提供创新产品和服务。药物筛选试剂盒可以分析拓扑异构酶活性药物,用于癌症化疗和抗菌方面。TopoGEN公司还能提供原核和真核化验拓扑异构酶 (I型,II,IV和旋转酶)检测试剂盒,体外体内诊断均可应用。TopoGEN公司还提供DNA载体、DNA裂解目标、拓扑异构酶抑制剂、拓扑异构酶抗体和相关试剂盒等完整产品线。

TopoGEN书-上海金畔

 

topogen产品目录:

货号

品名

规格

TG2000H-1

Human Topoisomerase IIa

250 units

TG2000H-2

Human Topoisomerase IIa

500 units

TG2000H-3

Human Topoisomerase IIa

1000 units

TG2000H-4

Human Topoisomerase IIa

2000 units

TG2005H-RC1

Human Topoisomerase I

250 units

TG2005H-RC2

Human Topoisomerase I

500 units

TG2005H-RC3

Human Topoisomerase I

3000 units

TG2005H-Y723F

Mutant Human Topoisomerase I (Y723F)

10ug

TG2000G-1

E. coli DNA Gyrase

100 units

TG2000G-3

E. coli DNA Gyrase

500 units

TG2000G-5

E. coli DNA Gyrase

1000 units

TG2000G-7

E. coli DNA Gyrase

2000 units

TG2000GSA-1

S. aureus DNA Gyrase

100 units

TG2000GSA-3

S. aureus DNA Gyrase

500 units

TG2000GSA-5

S. aureus DNA Gyrase

1000 units

TG2000EC-1

E. coli Topoisomerase IV

250 units

TG2000EC-3

E. coli Topoisomerase IV

500 units

TG2000EC-5

E. coli Topoisomerase IV

1000 units

TG1050-1

Topoisomerase Type IIa Nuclear Extraction/Assay Kit

100 assays

TG1001-1

Topoisomerase II Assay Kit (20ug kDNA)

100 assays

TG1001-2

Topoisomerase II Assay Kit (50ug kDNA)

250 assays

TG1001-1A

Topoisomerase II Assay Kit (20ug kDNA + 100u Human Topo Iia)

100 assays

TG1001-2A

Topoisomerase II Assay Kit (50ug kDNA + 250u  Human Topo Iia)

250 assays

TG1001-3A

Topoisomerase II Assay Kit (100ug kDNA + 500u Human Topo Iia)

500 assays

TG1009-1

Topoisomerase II Drug Screening Kit (25ug SC DNA)

100 assays

TG1009-2

Topoisomerase II Drug Screening Kit (62.5ug SC DNA)

250 assays

TG1009-1A

Topoisomerase II Drug Screening Kit (25ug SC DNA + 100u Topo Iia)

100 assays

TG1009-2A

Topoisomerase II Drug Screening Kit (62.5ug SC DNA + 250u Topo Iia)

250 assays

TG1009-3A

Topoisomerase II Drug Screening Kit (62.5ug SC DNA + 500u Topo Iia)

250 assays

TG1019-1

Topoisomerase II Drug Screening Kit (20ug kDNA)

100 assays

TG1019-2

Topoisomerase II Drug Screening Kit (50ug kDNA)

250 assays

TG1019-1A

Topoisomerase II Drug Screening Kit (20ug kDNA + 100u Human Topo Iia)

100 assays

TG1019-2A

Topoisomerase II Drug Screening Kit (50ug kDNA + 250u  Human Topo Iia)

250 assays

TG1019-3A

Topoisomerase II Drug Screening Kit (50ug kDNA + 500u  Human Topo Iia)

250 assays

TG1022

Topo II In Vivo Link Kit

200 assays

TG1007-1

E. coli Topoisomerase IV Drug Screening Kit (25ug SC DNA)

100 assays

TG1007-2

E. coli Topoisomerase IV Drug Screening Kit  (62.5ug SC DNA)

250 assays

TG1007-1A

E. coli Topoisomerase IV Drug Screening Kit (25ug SC DNA + 100u EC Topo IV)

100 assays

TG1007-2A

E. coli Topoisomerase IV Drug Screening Kit  (62.5ug SC DNA + 250u EC Topo IV)

250 assays

TG1007-3A

E. coli Topoisomerase IV Drug Screening Kit  (62.5ug SC DNA + 500u EC Topo IV)

250 assays

TG1008-1

E. coli Topo IV Drug Screening Kit (20ug kDNA)

100 assays

TG1008-2

E. coli Topo IV Drug Screening Kit (50ug kDNA)

250 assays

TG1008-1A

E. coli Topo IV Drug Screening Kit (20ug kDNA + 100u E. coli Topo IV)

100 assays

TG1008-2A

E. coli Topo IV Drug Screening Kit (50ug kDNA + 250u E. coli Topo IV)

250 assays

TG1008-3A

E. coli Topo IV Drug Screening Kit (50ug kDNA + 500u E. coli Topo IV)

250 assays

TG1040-1

Topoisomerase Type I Nuclear Extraction/Assay Kit

100 assays

TG1015-1

Topoisomerase I Assay Kit (25ug SC DNA)

100 assays

TG1015-2

Topoisomerase I Assay Kit (62.5ug SC DNA)

250 assays

TG1015-1A

Topoisomerase I Assay Kit (25ug SC DNA +100u Human Topo I)

100 assays

TG1015-2A

Topoisomerase I Assay Kit (62.5ug SC DNA + 250u Human Topo I)

250 assays

TG1015-3A

Topoisomerase I Assay Kit (125ug SC DNA + 500u Human Topo I)

500 assays

TG1018-1

Topoisomerase I Drug Screening Kit (25ug SC DNA)

100 assays

TG1018-2

Topoisomerase I Drug Screening Kit (62.5ug SC DNA)

250 assays

TG1018-1A

Topoisomerase I Drug Screening Kit (25ug SC DNA + 100u Human Topo I)

100 assays

TG1018-2A

Topoisomerase I Drug Screening Kit (62.5ug SC DNA + 250u Human Topo I)

250 assays

TG1018-3A

Topoisomerase I Drug Screening Kit (62.5ug SC DNA + 500u Human Topo I)

250 assays

TG1021

Topoisomerase I In Vivo Link Kit

200 assays

TG1017-1

Human Topoisomerase I High Load Kit

500 units

TG1017-2

Human Topoisomerase I High Load Kit

1000 units

TG1010-1

In Vitro SDS-K+ Precipitation Kit

100 assays

TG1010-2

In Vitro SDS-K+ Precipitation Kit

250 assays

TG1025

DNA Intercalator/Unwinding Kit

50 experiments

TG1011-1

In Vivo SDS-K+ Precipitation Kit

100 assays

TG1011-2

In Vivo SDS-K+ Precipitation Kit

250 assays

TG1020-0

Human Topoisomerase I ICE Assay Kit (No Antibody)

24 Tests

TG1020-1

Human Topoisomerase I ICE Assay Kit

24 Tests

TG1020-2a

Human Topoisomerase IIa ICE Assay Kit

24 Tests

TG1020-2b

Human Topoisomerase IIb ICE Assay Kit

24 Tests

TG1020-3a

Human Topoisomerase IIIa ICE Assay Kit

24 Tests

TG1020-3b

Human Topoisomerase IIIb ICE Assay Kit

24 Tests

TG1030-1

Human Topoisomerase I EMSA Kit

25 assays

TG1030-2

Human Topoisomerase II EMSA Kit

25 assays

TG1003

DNA Gyrase Assay Kit

100 assays

TG2000G-1KIT

Purified E. coli DNA Gyrase and Relaxed DNA Assay Kit (50ug Relaxed DNA + 100u Gyrase)

100 assays

TG2000G-3KIT

Purified E. coli DNA Gyrase and Relaxed DNA Assay Kit (250ug Relaxed DNA + 500u Gyrase)

500 assays

TG2000G-5KIT

Purified E. coli DNA Gyrase and Relaxed DNA Assay Kit (500ug Relaxed DNA + 1000u Gyrase)

1000 assays

TG2000G-7KIT

Purified E. coli DNA Gyrase and Relaxed DNA Assay Kit (1mg Relaxed DNA + 2000u Gyrase)

2000 assays

TG2001G-1KIT

Purified E. coli DNA Gyrase Drug Screening Kit (50ug Relaxed DNA + 100u Gyrase)

100 assays

TG2001G-3KIT

Purified E. coli DNA Gyrase Drug Screening Kit (250ug Relaxed DNA + 500u Gyrase)

500 assays

TG2001G-5KIT

Purified E. coli DNA Gyrase Drug Screening Kit (500ug Relaxed DNA + 1000u Gyrase)

1000 assays

TG2001G-7KIT

Purified E. coli DNA Gyrase Drug Screening Kit  (1mg Relaxed DNA + 2000u Gyrase)

2000 assays

TG2017-1

Mouse Monoclonal Antibody to Human Topoisomerase I (Covalent Complex)

100 units

TG2017-2

Mouse Monoclonal Antibody to Human Topoisomerase I (Covalent Complex)

250 units

TG2011-5

Mouse Monoclonal Antibody to Human Topo IIB

100 units

TG1024-A

Topoisomerase IIa Band Shift Kit

25 assays

TG1024-B

Topoisomerase IIb Band Shift Kit

25 assays

TG2010-1

Mouse Monoclonal Antibody to Human Topoisomerase IIα

50 units

TG2011-1

Rabbit Polyclonal Antibody to Human Topoisomerase IIα

250 units

TG2010-3

Rabbit Polyclonal Antibody to Human Topoisomerase IIb

250 units

TG2012

Human Antibody to Human Topoisomerase I (Scleroderma Serum)

500 units

TG2012-2

Rabbit Polyclonal Antibody to Human Topoisomerase I (Rabbit Serum)

250 ul

TG2012-4

Mouse Monoclonal Antibody to Human Topoisomerase I

100 ug

TG2014

Rabbit Polyclonal Antibody to Yeast Topo II (Serum)

250 ul

TG2010-4

Rabbit Polyclonal Antibody to Human Topoisomerase IIIb

250 units

TG2015

Mouse Monoclonal Antibody to Human Topoisomerase IIIα

50 ul

TG2011-2

Topoisomerase IIα C-terminal Peptide

50 ug

TG2011-3

Topoisomerase IIα MW Marker

20 units

TG2011-4

Topoisomerase I WB Marker

20 units

TG1030

Topoisomerse II Immunofluorescence Kit

>50 analyses

TG1035

Topoisomerase II Immunoprecipitation Kit

>50 analyses

TG1004-1

Human TDP1 Assay Kit  (Oligo)

100 assays

TG1004-1A

Human TDP1 Assay Kit  (DNA/* Complex)

100 assays

TG1004-2

Human TDP1 Assay Kit (Oligo)

250 assays

TG1004-2A

Human TDP1 Assay Kit (DNA/* Complex)

250 assays

TG2004H-1

Human TDP1 Protein

250 units

TG2004H-2

Human TDP1 Protein

500 units

TG2016

Rabbit Polyclonal Antibody to Human TDP1

250 units

TG2038-1

Human TDP2 DNA Substrate

10 ug

TG2038-3

Human TDP2 DNA Substrate

50 ug

TG1005-1

Human TDP2 Assay Kit

100 assays

TG1005-2

Human TDP2 Assay Kit

250 assays

TG2003H-1

Human TDP2 Protein

25 ug

TG2003H-2

Human TDP2 Protein

50 ug

TG2013-1

Kinetoplast DNA (catenated)

25 ug

TG2013-2

Kinetoplast DNA (catenated)

50 ug

TG2013-3

Kinetoplast DNA (catenated)

250 ug

TG2018-1

Linear Kinetoplast DNA Marker

10 ug

TG2020-1

Decatenated Kinetoplast DNA Marker

10 ug

TG2023-2

pRYG DNA (topo II target site)

20 ug

TG2025-1

Relaxed pRYG DNA Marker

10 ug

TG2028-1

Linear pRYG DNA Marker

10 ug

TG2030-1

Supercoiled pHOT-1 DNA

25 ug

TG2030-2

Supercoiled pHOT-1 DNA

50 ug

TG2030-3

Supercoiled pHOT-1 DNA

250 ug

TG2030-4

Supercoiled pHOT-1 DNA

500 ug

TG2035-1

Relaxed pHOT-1 DNA

25 ug

TG2035-2

Relaxed pHOT-1 DNA

50 ug

TG2035-3

Relaxed pHOT-1 DNA

250 ug

TG2035-4

Relaxed pHOT-1 DNA

500 ug

DR3000 HRHeLa-Sce

HR based Kit in HeLa Requiring I-Sce1 Transfection

ea

DR3000 HRHeLa-CPR

HR based Kit in HeLa Requiring CRISPR Transfection

ea

DR3000 HRCN1.4-Sce

HR based Kit in Mouse Neuronal Cells Requiring I-Sce1 Transfection

ea

DR3000 HRSH-Sce

HR in human Neuroblastoma Cells req. I-Sce1 Transfection

ea

DR3000 HR3T3-Sce

HR based Kit in MOUSE 3T3 Cells  Requiring I-Sce1 Transfection

ea

DR3000 Custom-Sce

Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.)  Includes HRHeLa cell line control

ea

DR3000A Custom-Sce

Customer makes own cell line hosting HR Reporter (I-Sce Transfectn req.) No HRHeLa cell line control

ea

DR5001 HNHela-Sce

NHEJ Based HeLa Requires I-Sce1 Transfection

ea

DR5000 HNSH-Sce

NHEJ Based in Human Neuroblastoma Line requires I-Sce1 transfection

ea

DR5000 HNCN1.4-Sce

NHEJ Based in Mouse Neuronal CN1.4 cells requires I-Sce1 transfection

ea

DR5000 HN3T3-Sce

NHEJ Based Mouse 3T3 cells Requires I-Sce1 Transfection

ea

DR5000 Custom-Sce

Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  Includes iHNHeLa cell line control

ea

DR5000A Custom-Sce

Customer creates cell line hosting NHEJ Reporter (I-Sce Transfectn req.)  NO iHNHeLa cell line control

ea

DR7001 LE-HE

HeLa Cell Reporters to detect DNA Methylation (DM) Effectors:  Hypomethylating and Hypermethylating Effectors

ea

DR7001 LE

HeLa Cell Reporters to detect DNA Hypomethylating Effectors

ea

DR7001 HE

HeLa Cell Reporters to detect DNA Hypermethylating Effectors

ea

TG4110

Camptothecin 10mM

0.25ml

TG4120

Doxorubicin 10mM

0.25ml

TG4130

Daunomycin 10mM

0.25ml

TG4140

Etoposide 10mM

0.25ml

TG4150

m-AMSA 10mM

0.25ml

TG4160

Genistein 10mM

0.25ml

TG4170

Ellipticine 10mM

0.25ml

TG4180

Ciproflaxin

50 ul

TG4100

Topotecan 10mM

0.25ml

TG4101

Irinotecan 10mM

0.25ml

TG4102

9-Nitro-20 (S) Camptothecin 10mM

0.25ml

TG4103

10-Hydroxy Camptothecin 10 mM

0.25ml

TG4104

9-amino-CPT 10mM (0.25ml)

0.25ml

TG4010

Topo I Assay Buffer 10x

1ml

TG4030

Gyrase Assay Buffer 5x

1ml

TG4032

5x Topoisomerase IV Assay Buffer (2-part)

1ml

TG4040

Topo II Assay Buffer (2 part)

1ml

TG4060

10% SDS

1ml

TG4065

Sarkosyl 20%

5ml

TG4037

5x Stop Buffer

1ml

TG4038

10x Gel Loading Buffer

1ml

TG4039

10x Proteinase K

0.5ml

TG4035

20mM ATP

0.5ml

TG4045

5X SA Gyrase Assay Buffer

1ml

TG4055

5X Potassium Glutamate

1ml

TG4090

Topoisomerase II Dilution Buffer

1ml

 

 

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TopoGEN TG2000G-1KIT说明书

 

 

TopoGEN 公司是一家专门为拓扑酶研究提供试剂和试剂盒。试剂盒可以有效地分析拓扑异构酶 I , II 和旋转酶,分析细胞内的拓扑抑制以及描绘拓扑异构酶抑制剂的特性。另外公司还提供 DNa c底物,拓扑抑制剂和抗体等。

 

TopoGEN TG2000G-1KIT说明书

 Purified E. coli DNA Gyrase and Relaxed DNA Kit (plasmid based). 
Catalog Numbe  TG2000G-1KIT        100 Reaction Set [50 ug DNA] 

Catalog Number TG2000G-3KIT         500 Reaction Set [250 ug DNA]

Catalog Number TG2000G-5KIT         1000 Reaction Set [500 ug DNA] 

Catalog Number TG2000G-7KIT          2000 Reaction Set [1 mg DNA]  
 
 
试剂的运输和储存试剂盒在干冰上运输。收到DNA后,应将其储存在4°C,并将缓冲器储存在-20°C。避免与质粒频繁的冻融循环,因为这可能导致DNA断裂。将酶储存在-70°C下。我们还建议在解冻后对酶进行校准(反复冷冻/解冻可能导致活性丧失);酶活性在冰上稳定1-3天。

 

一、导言
A.小结本试剂盒含有纯化细菌(大肠杆菌)DNA回旋酶,纯化至均一(基于SDS-PAGE)。DNA回旋酶是从高表达菌株制备的,在A2B2复合物中作为纯化的全酶供应。酶以试剂盒提供的上述质量控制数据中给出的单位浓度供应。DNA回旋酶储存在稳定缓冲液中[50 mm Tris-Cl pH 7.5,100 mm KCl,2 mm二硫三乙二醇,1 mm EDTA,50%甘油]。还包括在te中提供的底物dna(10 mm tris hcl,1 mm edta,ph 7.5)。松弛DNA的DNA浓度显示在试管的标签上。
DNA回旋酶检测试剂盒是基于对提供的松弛质粒DNA底物(pOR322的衍生物phot1)的超螺旋作用和大约2.7kb。在以下条件下,一单位回旋酶将在37℃下1小时内过冷500纳克(0.5微克)。反应机理如图1所示。该酶通过一个符号反转模型将负超螺旋引入松弛的phot1质粒底物中。能量共因子(atp)是*超驰反应所必需的。图1中的嵌入凝胶图像代表基于缺少溴化乙锭的琼脂糖凝胶的典型底物和产物(超螺旋DNA)结果。这些非eb凝胶是检测回旋酶活性的理想选择,这一点已被超螺旋和松弛dna形式之间的优异分辨率所证实。
图形

c.dna回旋酶的质量控制。1。通过检测线性运动细胞dna(kdna)和线性质粒dna的形成,进行核酸酶污染试验。1微克连环kdna或超螺旋puc19 dna的孵育(4小时。在37℃下,在10 mm mgcl2)存在下进行。在这些条件下不产生线性dna或分解产物。2。经SDS-PAGE分析,A和B亚基纯度均大于95%,且无内切酶。三。注:由于DNA回旋酶的高输入水平,由于自发流产反应,可能会检测到少量的线性DNA。这很正常。

d.纯化dna回旋酶的稀释缓冲液稀释应在50 mm tris-cl(ph 7.5)、100 mm nacl、2 mmdithiothriteol、1 mm edta和50%甘油中进行。

e.超卷绕试验条件一单位回旋酶通常与0.1至0.5微克松弛质粒DNA在20-30微升的反应体积中孵育1小时。在37摄氏度的缓冲液中。
AASSAY缓冲液(1份配方如下所示;试剂盒包括基于此配方的5份储备):35 mm Tris-Cl pH 7.5 24 mm KCl 4 mm MGCl2 2 2 mm二硫苏糖醇1.8 mm亚精胺1 mm ATP 6.5%甘油0.1 mg BSA/ml

f.放松dna质量控制试验:1。使用A260:A280读数用分光光度法评估纯度。2。在37摄氏度下单独用回旋酶缓冲液孵育60分钟,并没有形成刻痕或线性DNA物种。三。用拓扑根纯化的dna拓扑异构酶i(tg2005h-rc1)对超螺旋dna进行弛豫。在这些条件下,95%以上的质粒被放松。

 

g.试剂盒内容物。给出了TG2000G-1KIT尺寸的体积。(对于较大的套件尺寸,请根据需要乘以卷,请参见第1页了解套件尺寸)。

第1页规定了松弛的PHOT1 DNA(总共50微克)浓度。

超螺旋PYD1 DNA(25 UL凝胶负载缓冲液)。加载2 ul作为标记。

5倍回旋酶分析缓冲液(600微升)1倍缓冲液包含上述配方。

稀释缓冲液(600 ul)。用这个缓冲液稀释回旋酶。上述稀释缓冲液配方。

5X停止缓冲/凝胶负载染料(600 UL):5X缓冲液为5%肉桂基,0.125%溴酚蓝,25%甘油。

纯化DNA回旋酶(单位定义见第1页)。
h.典型反应混合物的方案(终体积为20 ul)

协议概要:反应体积应为20-30UL终体积(一般受可加载到凝胶上的体积的限制)。反应是在微乳管中与水、缓冲液和底物phot1松弛的dna组装的。后加入乙酰胆碱酯酶,反应在37℃孵育15~60分钟(或更长时间),然后用停止/负载缓冲液、蛋白酶K消化、提取和加载到琼脂糖凝胶上结束(注意,在电泳过程中不添加溴化乙锭(EB)到凝胶或凝胶缓冲液中。在电泳分离后,使用EB对凝胶进行染色。在某些情况下,您可能希望运行EB凝胶(凝胶和缓冲液中的0.5U/ML EB),以解决圆形基片(松弛光1)和超螺旋DNA产物的缺口和线状DNA。如果您不确定要运行哪个,我们建议您尝试运行两个凝胶系统,并将您的样品分为两个部分。通常在反应中有足够的DNA来运行两种凝胶体系。

i.样品反应(20 ul,显示添加顺序):

▪无菌蒸馏H20:根据需要变化,使体积达到20 ul▪5x分析缓冲液:4 u▪phot1松弛DNA:1 ul(根据需要变化)**松弛DNA的数量取决于灵敏度和检测方法。用溴化乙锭常规染色,约250纳克的DNA通常是足够的;然而,较少的可用于更敏感的染色。回旋酶:1 ul(始终在冰上后添加酶,并将所有试管转移到热块以启动反应序列)。

脱氧核糖核酸凝胶的非EB凝胶分析方法:
1。在37℃下孵育30-60分钟。

2。添加1/5体积的停止缓冲液/加载染料。
三。加入蛋白酶k至50ug/ml,37℃消化10-30min(可选步骤)。

4。加入20ul氯方:异戊醇(24:1混合物),短暂旋涡,抽出蓝色,水相。

5。将蓝色相装入1%琼脂糖(50x TAE缓冲液:242 g Tris碱、57.1 ml冰醋酸、100 ml 0.5m EDTA)。

6。电泳直到染料从凝胶中传播60-75%。

7。用0.5 ug/ml溴化乙锭染色30分钟(注意,EB是诱变剂,戴手套)。

8。去污(蒸馏水)室温10-30分钟(用手套处理凝胶)。

9。用紫外线透照仪拍摄。

电泳分析反应产物:对于每一种凝胶,进行阴性对照(松弛DNA缺乏促旋酶)。阴性对照组看起来与回旋酶反应产物非常不同(回旋酶反应产物迁移速度更快,形成超螺旋DNA;见图2)。

 

常见问题。

什么是关键的控制,使我可以清楚地识别药物,是影响DNA回旋酶使用这种分析?-标记dna(超螺旋、线性dna)是非常重要的。-一定要进行阳性药物控制(如氟喹诺酮类药物),以显示良好的切割活性。你应该看到线性DNA的增加。-包括一个阴性对照(要么没有药物,要么是topo ii药物,如依托泊苷)。未提供vp16或etoposide,但我们已提供。-一定要检查溶剂效应。二甲基亚砜或甲醇等溶剂用于溶解一些试验药物。用无药物但有溶剂(如1%二甲基亚砜)的对照反应进行试验。

我应该买哪种琼脂糖?任何无核酸酶琼脂糖的合理质量,从任何来源都可以使用(西格玛奥尔德里奇工程)。

什么是好的凝胶缓冲液使用?琼脂糖凝胶(1%)和流动缓冲液可以是任何标准的非变性电泳缓冲液(例如,制备50X的TAE凝胶缓冲液:242克TrI碱,57.1毫升冰醋酸和100毫升0.5米EDTA)。稀释到1X用于凝胶分离。确保凝胶也有1X的TAE缓冲液。
我应该使用EB凝胶还是非EB凝胶?如何使用EB凝胶?-通常,在不存在溴化乙锭(EB)的情况下,可以使用1%种凝胶(这些凝胶是测试酶活性的理想材料);然而,在该凝胶体系中,不能清楚地识别切口圆形的DNA产物。如上所述,含有EB的凝胶(0.5微克/毫升,EB在凝胶和缓冲液中)将提高裂解产物的分辨率(切口开环和线性DNA)。在对数据进行光聚焦之前,请务*水冲洗15分钟。-在某些情况下,取决于凝胶是如何运行的,拓扑异构体分布会干扰您看到卵裂产物的能力;然而,EB凝胶去除了这种并发症。-重要的是:如果不确定是否运行EB或非EB凝胶,我们建议您同时运行。简单地把你的反应分成相等的部分,同时运行两个凝胶。这样,你一定会看到所有的反应产物,并加强你对实验的解释。凝胶体系中的所有标记(例如,超螺旋、松弛、线性),将获得非常清楚和明确的结果。

时间和电压方面的运行条件是什么?-以1.5-2 V/cm(在电极之间测量)的速度运行凝胶,直到染料前沿移动约80%——运行后,非EB凝胶应在光照前用EB(0.5 ug/ml)染色15-30分钟,然后在水中或缓冲液中降解15分钟。EB凝胶在0.5μg/ml(凝胶和流动缓冲液)的存在下运行,然后在光记录之前用水染色15分钟。-重要提示:尽量不要让凝胶在一夜之间运行,但保持电泳时间少于1-2小时。长时间导致带扩散,降低凝胶效果的质量。

 

你建议这些分析的反应量是多少?-反应体积应为20-30UL终体积(受可加载到琼脂糖凝胶的威尔斯的体积的限制)。-这些反应应该在冰上的微乳管中进行(水、缓冲液、DNA、测试化合物和酶,应后添加)。-加入酶后,应将试管转移到加热块以启动反应。

不管我是用大规格的凝胶还是迷你凝胶都有关系吗?-两种都可以。微凝胶非常方便,运行时间也相当快。-井槽的实际形状和大小是影响波段分辨率的一个因素。好用一把长而薄的长方形梳子

终止条件对检测解理是否至关重要?-是的。反应应孵育30分钟(37°C),终止于快速加入1/10体积的10% SDS,然后在加载凝胶之前用50μg/ml蛋白酶K消化。将sds添加到37°的反应中,以便于将酶捕获在裂解复合物中。-此外,如果在加入sds之前加热、冷却或用高盐处理反应,topo断裂和再密封平衡可能会改变,断裂可以再密封。

为什么需要蛋白酶k?-捕获topo/dna复合物的药物会在dna和蛋白质(topo)之间诱导共价复合物,这种蛋白质必须被去除(降解)。不这样做将阻止解理产物的检测。-如果在sds之前对反应进行加热、冷却或高盐处理,topo-dna断裂和再密封平衡可能会改变,断裂可以再密封。

你能帮我们解释一下数据吗?-是的,我们一定能帮上忙!好的方法是将您的数据(support@topogen.com)和实验的完整描述发送给我们。我们会很快给你反馈。
你能给我们看一些真实的凝胶数据并讨论结果吗?-是的,我们可以。结果和有益的讨论如图2所示(见图例)。

 

图2.在非EB凝胶体系上分解反应产物

回旋酶反应在20ul的终体积中进行(见上述方案)。用1%sds终止反应,蛋白酶k消化,cia萃取。凝胶在50V下运行45-50分钟,并用EB(非EB凝胶)按上述方案染色。数据显示了松弛DNA的位置以及回旋反应产物(超螺旋DNA)。与氟喹诺酮(环丙沙星)的反应在右边后一条车道上产生一个线性的DNA切割产物(用一个小三角形标记)。对于凝胶数据的完整解释,重要的是包括放松的DNA、线性DNA标记以及控制旋回酶和回转酶+药物,如这里所示。