E. coli Topoisomerase IV
- E. coli Topoisomerase IV
- E. coli Topoisomerase IV Relaxation Kits
- E. coli Topoisomerase IV Decatenation Kits
- E. coli Topoisomerase IV Assay Kits for Cell Extracts
- E. coli Topoisomerase IV Cleavage Assay Kits
- High / Medium-Throughput Assay Kit - E. coli Topoisomerase IV
E. coli Topoisomerase IV
Topo IV (from Escherichia coli) is prepared by overexpressing the parC and parE subunits in E. coli and purifying them by methods adapted from Peng and Marians, 19991.
The parC and parE subunits are purified to >95% purity as judged by SDS-PAGE. The topo IV is supplied as a heterotetramer complex at a concentration of 10 U/µl in Dilution buffer.
It is recommended that the enzyme is aliquoted to avoid repeated freeze-thaw cycles. Store at -80ºC.
All enzyme is supplied with 5X concentrated Assay Buffer and Dilution buffers which are also available separately.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| T4001 | E. coli Topo IV | 100 U | £110 |
| T4002 | E. coli Topo IV | 500 U | £410 |
| T4003 | E. coli Topo IV | 1000 U | £820 |
| T4004 | E. coli Topo IV | 2000 U | £1540 |
E. coli Topoisomerase IV Relaxation Kits
These contain the enzyme and the supercoiled DNA substrate in addition to the Assay and Dilution buffers for relaxation reactions. 1 U of topo IV will relax 0.4 µg supercoiled pBR322 DNA in 30 minutes at 37°C.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| R4001 | E. coli Topo IV Relaxation Kit 1 | 100 U of E. coli Topo IV and 50 µg of Supercoiled DNA | £180 |
| R4002 | E. coli Topo IV Relaxation Kit 2 | 500 U of E. coli Topo IV and 250 µg of Supercoiled DNA | £670 |
| R4003 | E. coli Topo IV Relaxation Kit 3 | 1000 U of E. coli Topo IV and 500 µg of Supercoiled DNA | £1320 |
| R4004 | E. coli Topo IV Relaxation Kit 4 | 2000 U of E. coli Topo IV and 1 mg of Supercoiled DNA | £2480 |
E. coli Topoisomerase IV Decatenation Kits
These contain the enzyme and the kDNA substrate in addition to the Assay and Dilution buffers for decatenation reactions. 1 U of topo IV will decatenate 200 ng of kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl at 37°C for 30 minutes.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| D4001 | E. coli Topo IV Decatenation Kit 1 | 100 U of E. coli Topo IV and 20 µg kDNA | £180 |
| D4002 | E. coli Topo IV Decatenation Kit 2 | 500 U of E. coli Topo IV and 100 µg kDNA | £670 |
| D4003 | E. coli Topo IV Decatenation Kit 3 | 1000 U of E. coli Topo IV and 200 µg kDNA | £1320 |
| D4004 | E. coli Topo IV Decatenation Kit 4 | 2000 U of E. coli Topo IV and 400 µg kDNA | £2480 |
E. coli Topoisomerase IV Assay Kits for Cell Extracts
These kits are designed for assaying cell extracts and partially purified fractions containing over-expressed topo IV and contain supercoiled DNA substrate, Assay buffer, Dilution buffer, control relaxed DNA and stop buffer/loading dye.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| TE001 | E. coli Topo IV Assay Kit 1 for Cell Extracts | for 100 assays | £110 |
| TE002 | E. coli Topo IV Assay Kit 2 for Cell Extracts | for 500 assays | £350 |
| TE003 | E. coli Topo IV Assay Kit 3 for Cell Extracts | for 1000 assays | £690 |
| TE004 | E. coli Topo IV Assay Kit 4 for Cell Extracts | for 2000 assays | £1320 |
E. coli Topoisomerase IV Cleavage Assay Kits
These kits are designed specifically for cleavage reactions. They contain topo IV enzyme, supercoiled pBR322 DNA substrate and the Assay and Dilution buffers required for DNA cleavage reactions in addition to the enzyme and linearised pBR322 marker.
Cleavage specific enzyme available separately on request.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| TCK001 | E. coli Topo IV Cleavage Kit 1 | 100 U of E. coli Topo IV and 50 µg of supercoiled DNA | £POA |
High / Medium-Throughput Assay Kit - E. coli Topoisomerase IV
The kit is supplied with sufficient E. coli topo IV enzyme, plasmid DNA substrate, buffers and other assay components* for 100 assays. The enzyme is supplied at a concentration of 10 U/μl in Dilution Buffer. The kit is also supplied with sufficient wash buffers for one 96-well plate. These buffers are supplied as 20X concentrates and must be diluted with ultra pure water prior to use.
* Please note: The recommended fluorescence dye for this kit is SYBR Gold ® (Invitrogen) NOT SUPPLIED
More information about this assay can be found on the 'Services' page under 'High/Medium Throughput Assay'.
Kit issued with limited licence for individual use only.
Patent held by Plant Biosciences Ltd., Norwich, Norfolk, UK. ( Patent No. GB0424953.8)
| Cat No. | Product | Description | Price |
|---|---|---|---|
| TRIV01 | HTS Assay Kit - E. coli Topo IV 1-5 Kits | for 100 assays per plate | £365 |
| TRIV01 | HTS Assay Kit - E. coli Topo IV 6+ Kits | for 100 assays per plate | £285 |
References
- Peng, H. and Marians, K.J. (1999) Overexpression and purification of bacterial topoisomerase IV, in DNA Topoisomerase Protocols Vol. I (Bjornsti, M-A., and Osheroff, N. eds.), Humana Press, Totowa, N.Jersey pp.163-169
- Maxwell, A., Burton, N.P. and O'Hagan, N. (2006) High-throughput assays for DNA gyrase and other topoisomerases. Nucleic Acid Res. 34 (15), e104