Summary: Gel Shift, or Band Shift Assay, or Electrophoretic Mobility Shift Assay (EMSA) is a technique for studying gene regulation and determining protein:DNA interactions. The assay is based on the observation that complexes of protein and DNA migrate through a non-denaturing polyacrylamide gel more slowly than free DNA fragments or double-stranded oligonucleotides. The gel shift assay is carried out by first incubating a protein(s) (such as nuclear or cell extract) with a 32P end-labeled DNA fragment containing the putative protein binding site. The reaction products are then analyzed on a nondenaturing polyacrylamide gel. The specificity of the DNA-binding protein for the putative binding site is established by competition experiments using DNA fragments or oligonucleotides containing a binding site for the protein of interest or other unrelated DNA sequences.
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Protocols
- Gel Mobility Shift Assay for transcription factor binding (Contributed by Riddhish Shah, Hueley Lab)
Protocol can be used to investigate binding of transcription factor to your DNA seq. We use to see the differences due to polymorphic spot.
Added: Tue Apr 08 2003, Reviews: 0 Write review -
Band Shift Analysis
(Dario Neri's labs, Institute of Pharmaceutical Sciences ETHZ)
The procedure is for the determination of affinity constants and kinetic dissociation constants by band-shift assay refers to an ideal antibody fragment (e.g., a scFv or an Fab fragment) binding to a well-behaved protein antigen (pure, of well-defined oligomeric state, migrating as a single band in non-denaturing gel electrophoresis.
http://www.pharma.ethz.ch/institute_groups/biomacr...
Added: Mon Sep 09 2002, Hits: 3771, Reviews: 0 Write review Cached -
Band shifting
(Promega)
The gel shift assay is performed by incubating a purified protein, or a complex mixture of proteins (such as nuclear or cell extract preparations), with a 32P end-labeled DNA fragment containing the putative protein binding site. The reaction products are then analyzed on a nondenaturing polyacrylamide gel. The specificity of the DNA-binding protein for the putative binding site is established by competition experiments using DNA fragments or oligonucleotides containing a binding site for the protein of interest, or other unrelated DNA sequences.
http://www.promega.com/tbs/tb110/tb110.pdf
Added: Tue May 14 2002, Hits: 5457, Reviews: 0 Write review Cached -
EMSA
(Research Group of Johannes Schmid, Medical University of Vienna)
http://www.meduniwien.ac.at/user/johannes.schmid/E...
Added: Fri Nov 06 2009, Hits: 3503, Reviews: 0 Write review Cached -
EMSA Protocol
(Hammer Lab, University of Michigan)
Describes procedures including pouring Acrylamide Gel, Binding Reaction, Drying Gel and exposure
http://www.med.umich.edu/hammerlab/Protocols/EMSA%...
Added: Mon Mar 16 2009, Hits: 6317, Reviews: 0 Write review -
EMSA Using Oligos
(Mike A. Dyer)
Anneal two complementary oligos to make ds oligos as the probe for EMSA assay.
http://axon.med.harvard.edu/~cepko/protocol/mike/E...
Added: Tue May 14 2002, Hits: 2685, Reviews: 0 Write review Cached -
Gel Mobility Shift Analysis
(The Morimoto Lab, Northwestern University)
Precedures and recipes.
http://www.biochem.northwestern.edu/morimoto/resea...
Added: Mon Mar 29 2004, Hits: 4498, Reviews: 0 Write review -
Gel shift Assay
(The Hahn Lab, The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute, Seattle)
http://labs.fhcrc.org/hahn/Methods/biochem_meth/ge...
Added: Tue May 14 2002, Hits: 3942, Reviews: 0 Write review Cached -
Mobility Shift Assay
(Murphy Lab, Department of Pharmacology, Emory University)
http://www.pharm.emory.edu/tmurphy/Methods/mobilit...
Added: Tue Mar 03 2009, Hits: 2711, Reviews: 0 Write review -
Non-Radioactive EMSA Protocol
(Hammer Lab, University of Michigan)
Procedures include probe preparation by PCR amplifying a fragment of the promoter of interest using DIG-labeled dUTP in addition to normal dNTPs, Standard Nuclear Protein Extraction, Protein/Probe Binding, Electrophoresis, transfer and DIG detection
http://www.med.umich.edu/hammerlab/Protocols/Proto...
Added: Mon Mar 16 2009, Hits: 5162, Reviews: 0 Write review -
Radiolabeling of Probes for EMSA
(Mirmira Laboratory at the University of Virginia)
Labeling oligonucleotide using T4 kinase for gel shift assay.
http://faculty.virginia.edu/mirmira/resources_file...
Added: Sat Jun 05 2004, Hits: 2282, Reviews: 0 Write review Cached -
Super Shift Analysis
(The Morimoto Lab, Northwestern University)
Gel shift analysis with antibody
http://www.biochem.northwestern.edu/morimoto/resea...
Added: Mon Mar 29 2004, Hits: 2491, Reviews: 0 Write review