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. 2015 Sep 15;290(43):26033–26042. doi: 10.1074/jbc.M115.677286

FIGURE 2.

FIGURE 2.

TREM2-Ig precipitates ApoE from cerebrospinal fluid. A, TREM2-Ig, a human IgG1 isotype control antibody (hIgG), or unbound protein G beads were used as bait for immunoprecipitation of cynomolgus macaque CSF. Input (CSF), the TREM2-Ig regent by itself, and the precipitated products were separated by reducing SDS-PAGE and visualized by silver staining. TREM2-Ig, but neither hIgG nor beads alone, precipitated a doublet band of ∼36 kDa (white arrowhead), and a single band of ∼22 kDa (black arrowhead). Dashed lines indicate bands corresponding to TREM2-Ig, the 50-kDa IgG heavy chain (HC), and the 25-kDa IgG light chain (LC). B, immunoprecipitation (IP) of macaque CSF was repeated and analyzed by Western blot. The 36-kDa doublet was confirmed as ApoE and the 22-kDa single band as ApoA-I. ApoA-II, although not visible with silver staining, was detected by Western blot in both the CSF and TREM2-Ig precipitate lanes. The band indicated by the white arrowhead is the result of secondary antibody cross-reaction with the hIgG1 light chain. IB, immunoblot.