Expression and purification of AcLAPr and preparation of polyclonal antibody against AcLAPr.

(A) SDS-PAGE and Coomassie blue staining of the whole cell lysate corresponding to E. coli BL21 transformed with pCold-TF-AcLAP plasmid (lane 1), soluble fraction of the sonicated cell lysate (lane 2), eluted AcLAPr from the Ni-NTA agarose column (lane 3), thrombin-cleaved and purified AcLAPr (lane 4), and crude cyst extract at 72 h after induction of encystation of A. castellanii (lane 5). Protein marker (lane M). (B) The same samples as used for the experiment described in panel A were diluted 100-fold (lane 1–4), except crude cyst extract of A. castellanii (lane 5). Western blot analyses were carried out with rat polyclonal anti-AcLAPr antibody. (PDF)