Journal: Molecular & Cellular Proteomics : MCP
Article Title: Functional Characterization and Comparison of Plasmodium falciparum Proteins as Targets of Transmission-blocking Antibodies *
Figure Lengend Snippet: Heterologous expression of surface and secreted P. falciparum sexual-stage proteins. A , Coomassie brilliant blue R-250 staining of recombinant proteins resolved by reducing SDS-PAGE. Each protein has a C-terminal BAP and 6HIS and was purified from whole HEK293E cell supernatant by HisTrap Excel Ni-Sepharose column prior to visualization. B , Western blotting detection of hexahistidine tag affinity-purified proteins, which were resolved by reducing SDS-page, blotted, and probed with monoclonal anti-pentahistidine antibody. Each protein is shown with and without treatment by PNGaseF, an amidase which cleaves N -linked glycan residues. C , Western blotting detection of affinity-purified Pfs25 by 4B7 monoclonal antibody after SDS-PAGE under reducing and non-reducing conditions and nitrocellulose transfer.
Article Snippet: Recombinant proteins were resolved by 4–20% Tris-Glycine gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions and visualized by Coomassie brilliant blue R-250 (Bio-Rad, UK).
Techniques: Expressing, Staining, Recombinant, SDS Page, Purification, Western Blot, Affinity Purification