Skip to main content
. 2014 Jan 23;9(1):e87240. doi: 10.1371/journal.pone.0087240

Figure 1. Biochemical characterization of FLH.

Figure 1

(A). TEM of FLH negatively stained. The low-magnification micrograph of the purified protein shows the top (circles) and lateral (rectangles) views of the molecules. The insert shows a high-magnification view of the FLH molecules. (B). SDS-PAGE analysis under non-reducing conditions on a 3–7% polyacrylamide gradient gel stained with Coomassie blue. FLH shows one polypeptide in contrast to CCH and KLH that possess two (arrows). As controls, FCH and FMH were included. (C). SDS-PAGE analysis under reducing conditions on a 3–12% gradient polyacrylamide gel. The two CCH and KLH subunits are noted in contrast to the one subunit of FLH. (D). Native gel agarose electrophoresis. The image shows a single compacted band for each hemocyanin. (E). Chromatography of dissociated FLH. The previously dissociated protein was subjected to IEX HPLC on a MonoQ 5/5 column and eluted with a linear NaCl gradient (0 to 70%, green line). The chromatography was monitored at 280 nm (blue line). (F). SDS-PAGE analysis under reducing conditions of the elution fractions from the chromatography of E. The presence of a single main polypeptide in FLH was confirmed (G). FLH oligosaccharide pattern. Hemocyanins were blotted onto a nitrocellulose membrane, incubated with biotinylated lectins ConA and PNA and developed with avidin-FAL followed by NBT/BCIP detection.