Knight:Electrophoretic mobility shift assay: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
(No difference)
|
Revision as of 10:17, 25 August 2006
In progress! Has not been tested.
Overview
An assay to check for protein-DNA binding.
Materials
- SYBR Green EMSA nucleic acid gel stain
- 10,000X concentrate in dimethylsulfoxide
- Stored at -20°C
- SYPRO Ruby EMSA protein gel stain
- Light sensitive
- Store at room temperature
- Trichloroacetic acid (TCA)
- Loading solution
- Tom thinks we could get away with using our typical gel loading buffer.
- Binding buffer
- Need to determine composition. See Reshma Shetty/Scratchpad#DNA binding reaction conditions for notes.
Procedure
References
-
EMSA kit from Invitrogen
- Jing D, Beechem JM, and Patton WF. The utility of a two-color fluorescence electrophoretic mobility shift assay procedure for the analysis of DNA replication complexes. Electrophoresis. 2004 Aug;25(15):2439-46. DOI:10.1002/elps.200405994 |
- Jing D, Agnew J, Patton WF, Hendrickson J, and Beechem JM. A sensitive two-color electrophoretic mobility shift assay for detecting both nucleic acids and protein in gels. Proteomics. 2003 Jul;3(7):1172-80. DOI:10.1002/pmic.200300438 |
