Dear RG experts,
let me know any other technique
DamID can be used to mark protein-DNA interactions in-vivo, yet with the disadvantage that it can only be done in cultured cells and that the resolution is slightly lower than ChIP-seq (in the order of 1-2 kb).
See e.g.: http://www.ncbi.nlm.nih.gov/pubmed/18463634
Electrophoretic mobility shift assay (EMSA), also known as gel shift assay, can be used to detect protein-DNA interaction.
Fluorescence polarization with synthetic fluorescently-labeled oligonucleotides is another way.
Immunoblotting are also used for studying Protein-Dna interaction experimentally
Dear Vijey,
you can incubate your DNA with the protein in question and probe for your protein with an antibody that should bind where the DNA is bound , the the run an SDS PAGE gel.
If you have no binding of the antibody onto the protein, that means that your protein is bound with the DNA and you should see no band on your gel when compared with an unbound protein.
That my be a way of looking into it.
Best wish,
Nikos
This is a rather dated reference, but may be helpful depending on your application:
In vivo and in vitro characterization of protein interactions with the dyad G-box of the Arabidopsis Adh gene.W L McKendree, A L Paul, A J DeLisle, R J Ferl
The link and handbook may be helpful to evaluate the methods for detecting protein-DNA interactions
https://www.lifetechnologies.com/tr/en/home/life-science/protein-biology/protein-biology-learning-center/protein-biology-resource-library/pierce-protein-methods/methods-detecting-protein-dna-interactions.html
https://tools.lifetechnologies.com/content/sfs/brochures/1601945-Protein-Interactions-Handbook.pdf
- Badges
- Science method
- Similar topics
- Chemistry
- General Biochemistry
- Biochemical Methods