HMW-MAA-specific Ab response determined by FACS and IHC.

<p>(<b>A</b>) The human melanoma cell lines 518A2 and M14 were each incubated with 1 µg mAb VT80.12 (as positive control to detect HMW-MAA) or 100 µg purified rabbit IgG from immunizations with 15/3/6-KLH conjugate or KLH. Bound IgG were detected with FITC-labeled goat anti-mouse IgG or FITC-labeled goat anti-rabbit IgG. Histogram overlays show unstained (white) vs. stained cells (grey). (<b>B</b>) IHC on 518A2 tumor tissues of C.B.17 SCID/SCID mice <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019383#pone.0019383-Wagner2" target="_blank">[13]</a>. HMW-MAA staining was done with PBS buffer (negative control), mAb VT80.12 (positive control), and biotinylated IgG purified from rabbits immunized with 15/3/6-KLH conjugate (anti-15/3/6) or KLH (anti-KLH). Bound Abs were detected with StrepAB/HRP and visualized by DAB-chromogen solution and subsequent hematoxylin counterstaining.