Dilution PLIMSTEX

The work flow for dPLIMSTEX starts with half the volume of an equilibrated protein-peptide complex for measurement. The other half is diluted in aqueous buffer before incubation (Figure 11.5). The dilution step is continued until the concentration of the peptide is too low to be detected. dPLIMSTEX was first demonstrated by using a model system, calcium-saturated calmodulin with the opioid peptide )-endorphin it yielded a similar binding constant as that determined by standard PLIMSTEX and other methods. dPLIMSTEX was then applied to a monoclonal antinitro-tyrosine antibody, in complex with a 3-nitrotyrosine-modified peptide system. A binding stoichiometry of 1 2 was confirmed. In addition, a in the low nM range and a minimum of five amino... 

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