Western blot analysis of serum from chinchillas inoculated with the WT M. catarrhalis strain O35E.
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Equivalent amounts of whole cell lysates (WT M. catarrhalis O35E, uspA2 KO strain O35E.2, hag transposon mutant strain O35E.TN2, and ompCD KO strain O35E.CD1) were resolved by SDS-PAGE, transferred to PVDF and probed with the indicated primary Abs. Panels A and B: Pre- and post-infection serum samples were pooled and used as primary Abs at a dilution of 1∶250. Goat α-rat IgG conjugated to HRP were used as secondary Abs. Controls: The murine monoclonal Abs 10F3 (Panel C, α-CopB), 5D2 (Panel D, α-Hag), 17H4 (Panel E, α-UspA2) and 1D3 (Panel F, α-OMPCD) were used as primary Abs in combination with goat α-mouse HRP-(IgG+IgA+IgM) secondary Abs. These controls were included to verify the identity of proteins recognized by post-infection chinchilla serum in panel B. MW markers are shown to the left of in kDa.