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. 2019 Aug 30;294(43):15698–15710. doi: 10.1074/jbc.RA119.009980

Figure 4.

Figure 4.

V. cholerae TcpB binds to CTXφ pIII in ELISAs. A, CTXφ pIII protein was immobilized on ELISA microplate wells, and V. cholerae TcpB was added in increasing amounts in solution. Captured TcpB was detected with rabbit anti-TcpB(64–78) antibody and enzyme-linked secondary antibody. TcpB was immobilized as a positive control; negative controls were blocked wells lacking immobilized antigen (−) and immobilized antigens TcpA and BSA to show the specificity of the TcpB:pIII interaction. B, TcpB was immobilized on ELISA microplate wells, and pIII was added in increasing amounts in solution. Captured pIII was detected with anti-pIII antibody and enzyme-linked secondary antibody. pIII was immobilized as a positive control, and TcpA and BSA were immobilized as negative controls for capture. C, TcpB was immobilized, and pIII was added in increasing amounts in solution to inhibit anti-TcpB(64–78) antibody binding. Bound antibody was detected with enzyme-linked secondary antibody. TcpA and BSA were added in solution as negative controls for inhibition. For all experiments, absorbance was read at 405 nm. Values represents the average of three separate experiments; error bars represent the S.E.