Probably the easiest and most sensitive protein assay if you have access to a fluorescence microplate reader is our NanoOrange protein assay. http://www.probes.com/media/pis/mp06666.pdf This is a lipophilic dye that becomes fluorescent only in the SDS coating around the protein. You would add the reagent, heat to denature the protein and cool to measure the total protein. The signal is fairly independent of the nature of the protein and the sensitivity is as good as 10 ng/mL, which is about 50-100 times better than the Bradford, Lowry or BCA methods. Nucleic acids do not interfere with the NanoOrange assay. If your amount of immobilized protein is less than this amount then I don't know of any way to measure it, except if it was radioactively labeled. mquagr1 wrote: > Dear all, > > I am assaying prtein-protein interaction by ELISA and Iwould like to know if > there is a way I can ensure equal immobilization of my proteins to the > microtitre wells. For example, if there is a very sensitive assay that I can > use to measure the amount of protein immobilized so I know how much protein is > indeed present in the wells. Any ideas or suggestions? > > M.Quagraine
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