Additional file 1.
Immunofluorescence assay to demonstrate that monoclonal antibodies do not bind protein A in transfected T. cruzi epimastigotes expressing TcCLC fused with protein A/C. A) As a control, T. cruzi transfected epimastigotes were first incubated with Protein A antibody. Positive immunolabeling was observed at the cell anterior, in close proximity to the kinetoplast. B) Transfected epimastigotes incubated with a monoclonal antibody against TcCruzipain , which specifically recognizes the T. cruzi reservosomes at the cell posterior (arrow). C) Transfected epimastigotes incubated with a monoclonal antibody against TcNup , which specifically recognizes a protein associated to the nuclear membrane of T. cruzi. As expected, monoclonal antibodies in B and C were specific for their target proteins, but did not recognize CLC-A/C at the cell anterior. These results show that monoclonal antibodies do not cross react with the protein A/C tag in our transfected epimastigotes. Scale bar = 5 μm.
Format: TIFF Size: 2.2MB Download file
Kalb et al. BMC Cell Biology 2014 15:23 doi:10.1186/1471-2121-15-23