Figure 1.

Monoclonal antibody 17A12 detects an unknown protein (UP1) present only in M. avium subsp. paratuberculosis whole cell extracts. Shown are immunoblots of mycobacterial whole cell extracts exposed to MAbs. (A) The top blot was exposed to 17A12, which detects only the three MAP strains present in lanes 4, 9, and 11. The lower blot, labeled internal control, was exposed to MAb 4B6, which detects an unknown but highly conserved mycobacterial protein (Bannantine et al., 2007b) and shows the relative amounts of protein loaded in each of those lanes. The mycobacterial whole cell antigen prep used is indicated. Abbreviations: M, M. avium subsp. silvaticum; M. scro, M. scrofulaceum; M. abs, M. abcessus; M. ap, M. avium subsp. paratuberculosis; M. aa, M. avium subsp. avium; M. intracel, M. intracellulare. (B) UP1 is not present in M. avium subsp. avium or M. avium subsp. hominissuis isolates. The control blot labeled MMP in each image was exposed to a MAb previously developed in our laboratory that binds to the major membrane protein, which is present in all MAC species (Bannantine et al., 2007a). The upper blots are loaded with M. avium subsp. avium isolates and the lower blots are loaded with M. avium subsp. hominissuis isolates not analyzed in (A) (see Table A1 in Appendix for these strains). (C) Quantitative densitometry was performed on several MAP strains and the M. avium subsp. hominissuis strain 104. The results are expressed as a percent of 17A12 divided by the internal control (MMP). Error bars indicate standard deviations of the means. Data are representative of three independent culture replicates and error bars are standard deviation of the mean.