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. Author manuscript; available in PMC: 2012 Jun 18.
Published in final edited form as: J Neural Transm (Vienna). 2011 May 7;118(5):795–808. doi: 10.1007/s00702-011-0653-2

Figure 2. Lrrk2 reactivity in human peripheral blood mononuclear cells and isolated monocytes.

Figure 2

(A-B) Lrrk2 reactivity was detected in peripheral blood mononuclear cells (PBMC) (A) and anti-CD14+ monocytes (B) that were isolated by Ficoll separation and magnetic beads (carrying anti-CD14 antibody), respectively, from healthy adult controls (n=3) (Vranjkovic et al. 2011). FACS sorting of fixed and permeabilized cells was carried out using monoclonal anti-Lrrk2 antibody MJFF-1(C5-8). In each panel, a selection of cells is made by the forward and side scatter light properties (horizontal and vertical axis respectively in the left diagram) and a histogram (left diagram, horizontal and vertical axis represent the number of the cells counted and fluorescence respectively) of the fluorescence (IgG) profile of the Lrrk2 is generated to show the degree of the antibody binding. Red vertical lines show mean fluoresce of unstained cells. PBMC and monocytes were collected from two different individuals. Note, only 0.58% of PBMC (A) and 0.81% of monocytes (B) were gated in the absence of anti-Lrrk2 antibody. However, following the optimization of primary and secondary antibody concentrations the percentage of Lrrk2-positive cells measured 19.04 (A2), 24.54 (A3), and 25.71 (A4) for PBMC; and 1.8 (B2), 10.31 (B3), and 43.17 (B4). Thus, staining with anti-Lrrk2 revealed that <25.71% of all PBMC (A) and <43.17% of all monocytes (B) were successfully gated.