Skip to main content
. 2012 Apr 2;109(16):6229–6234. doi: 10.1073/pnas.1200470109

Fig. 2.

Fig. 2.

Identification of NSP4 specific rat monoclonal antibodies. (A) Only 3 of 14 monoclonal antibodies obtained from hybridomas after immunization of rats, named mAb 1C3, 2F6, and 8H3, were specific for NSP4. NSP4, the human Gzms (A, B, K, M, and H), and all known human NSPs were coated on microtiter plates. Supernatants of hybridoma clones secreting rat antibodies were analyzed in an ELISA using a secondary anti-rat HRP-conjugated polyclonal Ab. Percent reactivity was calculated by setting the mean reactivity of mAbs with NSP4 to 100%. Data are the mean from three independent experiments (n = 3, ± SD). (B) NSP4, the five human Gzms and human NSPs were separated by SDS/PAGE, transferred to Hybond ECL transfer membrane, and then probed with rat anti-NSP4 mAb 1C3, 2F6, or 8H3 and secondary HRP-labeled Ab. (C) Similar amounts of all tested proteins used as in A and B are shown after reducing SDS/PAGE and subsequent silver-staining. A horizontal arrow points to the glycosylated NSP4, carrying an amino-terminal extension (i.e., S-tag and an enterokinase cleavage peptide sequence).