Fig. 5
From: TREM2 splice isoforms generate soluble TREM2 species that disrupt long-term potentiation
Detection of TREM2 alternative isoforms protein expression in the human brain. A Design of customized antibodies to specifically detect human TREM2222 (Ab222) and TREM2219 (Ab219). B, C Western blot of TREM2222 and TREM2219 in cell extracts from HEK-293T transfected with tagged TREM2 isoforms (TREM2230, TREM2222, and TREM2219) using Ab222 (B), Ab219 (C), anti-human TREM2, and anti-β-actin antibodies. D Western blot analysis of TREM2219 expression in RIPA (upper blot) and TBS-soluble extracts (bottom blot) from the brain tissue of control human subjects (CTRL) and patients with Alzheimer’s disease (AD) using Ab219 and vinculin as a loading control. E ELISA of lysates from HMC3 cells transfected with TREM2 isoforms or empty plasmid (n = 3). The capture antibodies used were MAB17291 (N-terminal), Ab222, and Ab219. Data are shown as mean values ± SEM of optical density at 450 nm (O.D. 450 nm). F ELISA of purified soluble TREM2 proteins (10 ng/ml) using a TREM2 N-terminal capture antibody (MAB17291), as well as Ab222 and Ab219 to assess their specificity. Data are shown as O.D. 450 nm. G Concentration of TREM2222 and TREM2219 in a pool of human brain lysate determined by ELISA