Maltose binding protein (MBP) sandwich ELISA with immobilized malE-A1(Cys)
This application involves the identification of antigens in solution by sandwich ELISA using nanoCLAMPs covalently conjugated to maleimide coated microtiter wells via their lone C-terminal Cys. Antigen bound by the immobilized nanoCLAMP is identified by an enzyme-conjugated secondary antibody followed by development with the appropriate secondary substrate.
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Analysis of the interaction between malE-A1(Cys) and MBP by sandwich ELISA. malE-A1(Cys) was conjugated to maleimide coated microtiter wells of 96 well strips via the lone C-terminal Cys, at 5 µg/ml. Dilutions of biotinylated-MBP were incubated on the strips, washed, and then incubated with Streptavidin conjugated HRP and developed with TMB Ultra. Reactions were stopped with H2S04 and absorbance measured at A450. The blank wells were processed the same, except they included no malE-A1(Cys).