EMSA (Gel-Shift) Kits (CA - CF)
EMSA kits starting with CA - CF
EMSA (Electrophoretic Mobility Shift Assay) Kits are useful tools for validating results from Protein/DNA Arrays or identifying proteins that interact with DNA. You can also use them to assess the DNA-binding activity of a specific transcription factor. This rapid technique is based on the different electrophoretic mobilities of free DNA and protein/DNA complexes in native (non-denaturing) polyacrylamide gels.
Here’s how it works
- Incubate the biotin-labeled DNA probe with your nuclear extracts which should contain your protein(s) of interest.
- Separate the mixture on a nondenaturing polyacrylamide gel.
- Transfer the protein/DNA complex to a nylon membrane. Incubate the blot with streptavidin-HRP and then add chemiluminescent substrate.
- Visualize shifted bands using film or imaging system that correspond to the protein DNA complexes by chemiluminescence.
To confirm whether the DNA-binding activity of a particular protein is specific, you simply add an excess of cold, unlabeled DNA probe to the reaction mixture. If binding is specific, the cold probe competes with the labeled DNA probe for binding to the protein. As a result, the shifted band is eliminated.
For some Transcription Factors, multiple consensus sequences for the same TF have been published in the literature and, where appropriate, we have provided those variations available in our EMSA kits and arrays. For the TF’s with multiple consensus sequences, they have been denoted with numbers in parenthesis, i.e. (1), (2), (3) etc. Also, PR = Promoter, BP = Binding Protein. To obtain the consensus sequences and references for the TF’s, used in our arrays, please contact our Technical Support group.
Biotin-labeled and cold TF probes
Biotin-labeled and cold control probes
Control nuclear extract
Each EMSA Kit is supplied with enough reagents to run 25 reactions.
To prepare nuclear extracts for this procedure, please see the Nuclear Extraction Kit, PN AY2002.
Custom EMSA kit creation
If you can’t find your EMSA on our list or website, we can custom manufacture your EMSA kit. Just provide the sequence or TF name and we will provide all of the components supplied in all of our kits.
EMSA probe sets
In addition to our EMSA Kits, we offer individual EMSA probes for purchase without the other EMSA Kit components. USB EMSA TF Probe Sets are intended for use with our EMSA Kits. Each TF probe set is provided as 25 µl (10 ng/µl) of biotin-labeled TF probe accompanied by 25 µl (330 ng/µl) of cold probe, its unlabeled form.
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