Mobility shift assay protocols can be easily converted to infrared fluorescent assays by replacing the existing DNA oligonucleotides with IRDye® Infrared Dye end-labeled oligonucleotides. Binding and electrophoresis conditions are the same as any other EMSA detection method.
A DNA oligonucleotide end-labeled with IRDye 700 infrared dye is a good substrate for protein binding. Using the Odyssey® Imager, IRDye Infrared Dye labeled DNA detection is linear within a 50-fold dilution range from 9.1 fmol to 0.18 fmol.
Infrared fluorescent assays can be completed in less than two hours with no gel transfer or film exposure. The gel doesn’t even have to be removed from the glass plates for imaging. If you are not satisfied that the electrophoresis has progressed far enough, you can place the gel back into the electrophoresis unit and run longer.
Other IRDye 700 oligonucleotides as well as IRDye 800 oligonucleotides are available through Integrated DNA Technologies, TriLink BioTechnologies, or Metabion International AG.
| Infrared | Radioisotope |
|---|---|
| Easy access and disposal | Short half-life of the label |
| Dye is stable for a long time | Regulatory procedures Disposal limitations |
| Non-hazardous | Hazardous |
| Gel (glass plates) can be easily imaged on the Odyssey | Lengthy incubations with autoradiographic film |
| Detection of the probe is rapid | Time-consuming and inconvenient |
| Gel can be replaced back and run longer | Not possible |
Special pricing is available in your location. For more information, please contact your local expert here.
| Product Name | Price |
|---|---|
10X Orange Loading Dye
10X Orange Loading Dye is used as a loading buffer for DNA gel electrophoresis. This loading dye is not visualized when imaged on Odyssey Imagers. |
|
IRDye® 700 AP-1 Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying AP-1. |
|
IRDye® 700 CREB Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding of CREB. |
|
IRDye® 700 HIF-1 Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying HIF-1. |
|
IRDye® 700 NFĸB Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying NFĸB. |
|
IRDye® 700 p53 Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying p53. |
|
IRDye® 700 Sp-1 Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying SP-1. |
|
IRDye® 700 STAT3 Consensus Oligonucleotide
Use this IRDye labeled oligonucleotide in gel shift assays to assess protein binding when studying STAT3. |
|
Odyssey® EMSA Kit
The Odyssey EMSA Kit, coupled with IRDye 700 Oligonucleotides, is an excellent alternative method to radioisotopic and chemiluminescent EMSA detection methods. |