Electrophoretic mobility shift assay: Difference between revisions
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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 <sup>32</sup>P 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. | |||
==References== | |||
===Background information=== | |||
*[http://www.piercenet.com/Products/Browse.cfm?fldID=99A16E15-3470-4FD1-9A3C-0BBABBBE45F9 EMSA] by Pierce | |||
*[[Wikipedia:Electrophoretic mobility shift assay|Electrophoretic mobility shift assay]] from Wikipedia | |||
== | ===Protocols=== | ||
*[http://www.mrw.interscience.wiley.com/cp/cpmb/articles/mb1202/frame.html Mobility Shift DNA-Binding Assay Using Gel Electrophoresis] from Current Protocols in Molecular Biology | *[http://www.mrw.interscience.wiley.com/cp/cpmb/articles/mb1202/frame.html Mobility Shift DNA-Binding Assay Using Gel Electrophoresis] from Current Protocols in Molecular Biology | ||
*[http://www.molecularcloning.com/members/protocol.jsp?pronumber=2&chpnumber=17 Gel Retardation Assays for DNA-binding Proteins] from [[Molecular Cloning]] | *[http://www.molecularcloning.com/members/protocol.jsp?pronumber=2&chpnumber=17 Gel Retardation Assays for DNA-binding Proteins] from [[Molecular Cloning]] | ||
*[http://probes.invitrogen.com/media/pis/mp33075.pdf EMSA kit] from Invitrogen (primary advantage is that this protocol doesn't require use of radioactivity) | *[http://probes.invitrogen.com/media/pis/mp33075.pdf EMSA kit] from Invitrogen (primary advantage is that this protocol doesn't require use of radioactivity) | ||
Revision as of 12:21, 11 August 2006
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.
References
Background information
- EMSA by Pierce
- Electrophoretic mobility shift assay from Wikipedia
Protocols
- Mobility Shift DNA-Binding Assay Using Gel Electrophoresis from Current Protocols in Molecular Biology
- Gel Retardation Assays for DNA-binding Proteins from Molecular Cloning
- EMSA kit from Invitrogen (primary advantage is that this protocol doesn't require use of radioactivity)
