We are looking at whether a putative drug binds to a nuclear receptor? What experiments can be suggested? We have looked at possible interaction of the drug with the receptor using autodock and other softwares and have got several possible sites that the drug may bind to? We would like to do wet lab validation but dont have expertise in this area. Any suggestions on protocols and articles?
Thanks Dr. Mohammed
Can you please recommend any specific assay you may have used before for nuclear receptor from Life Technologies?
I am sorry if my question was vague but I dont have a lot of insight into this field.
Try to use marker to ligand. Radio Assay, immunoassay or fluorescent assay. Do you have something compound that fact binds to this region? If you have you can measure the connection, you just need to make a comparison. If not, you can try a labeled antibody to a drug and measure with a secondary antibody that connection.
Tag the drug with a fluorescent marker and then get it bound to the desired cells and watch through fluorescent microscope
If you want to see the interaction in-vitro then mass spectroscopy is the best. You can incubate pure protein (NHR) and you drug of interest at 37 degree C for different time points..then run SDS gel..cut the band and go for MALDI mass spectra analysis. you will get two peaks, one at NHR's MW and other at (NHR+drug) MW position.
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