Electrophoretic mobility shift assay
This assay permits testing of specific binding of proteins to DNA fragments. DNA that is bound to protein will migrate slower in a nondenaturing polyacrylamide gel than unbound DNA during electrophoresis.
Most protocols rely on 32P labelling of the DNA fragment. However, it is also possible to detect the DNA via nonradioactive detection methods (like fluorescence). Ethidium bromide staining is generally not sensitive enough since usually small amounts of DNA are used in this assay.
- Jing D, Beechem JM, and Patton WF. . pmid:15300760.
- Jing D, Agnew J, Patton WF, Hendrickson J, and Beechem JM. . pmid:12872218.
- Mobility Shift DNA-Binding Assay Using Gel Electrophoresis from Current Protocols in Molecular Biology
- Gel Retardation Assays for DNA-binding Proteins from Molecular Cloning (subscription required)
- EMSA kit from Invitrogen
- LightShift Chemiluminescent EMSA Kit from Pierce
- claimed to be more sensitive than radioactive and digoxigenin methods