Electrophoretic mobility shift assay

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*[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 or prelabelling of DNA, sensitivity is questionable though)
*[http://probes.invitrogen.com/media/pis/mp33075.pdf EMSA kit] from Invitrogen (primary advantage is that this protocol doesn't require use of radioactivity or prelabelling of DNA, sensitivity is questionable though)
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*[http://www.piercenet.com/Objects/View.cfm?type=ProductFamily&ID=06010701 LightShift Chemiluminescent EMSA Kit from Pierce] (claimed to be more sensitive than radioactive and digoxigenin methods)
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*[http://www.piercenet.com/Objects/View.cfm?type=ProductFamily&ID=06010701 LightShift Chemiluminescent EMSA Kit] from Pierce (claimed to be more sensitive than radioactive and digoxigenin methods)

Revision as of 18:43, 22 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

Protocols

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