Recombinant bovine CyPA (rbCyPA) is generated in an
A pbCyPA-PAL7 (6363 bp) was constructed to express rbCyPA in E.coli (BL21). B SDS-PAGE analysis showed that Profinity eXact-tagged rbCyPA was expressed when BL21 cells were treated with IPTG and obtained as a soluble protein. Tag-free rbCyPA was collected after affinity and gel chromatography. Lane 1: molecular weight marker, 2: Cell lysate of BL21 cells cultured without IPTG, 3: Cell lysate of BL21 cultured with IPTG, 4: Soluble proteins obtained by sonication of cell lysate of BL21 cultured with IPTG, 5: rbCyPA collected after affinity and gel chromatography. C Western-blot analysis was conducted using the commercial anti-CyPA antibody (1) or the control rabbit immunoglobulin (2) to confirm that the band shown on lane 5 of Figure 2B was bovine CyPA. D Neutralization with rbCyPA completely inhibited the immunoreactivity of anti-CyPA antibody used for immunohistochemistry. Scale bars = 100 μm.