6e10 monoclonal antibody

Synchronized L1 worms were cultured for 36 h at 16°C on 100 mm NGM plates containing OP50 and drugs, upshifted to 23°C, and incubated at that temperature for another 32 h. The worms were then collected in 1x phosphate-buffered saline (PBS) (8 g NaCl, 0.2 g KCl, 1.44 g Na₂HPO₄, and 0.24 g KH₂PO₄). The total protein was isolated with ice-cold radioimmunoprecipitation assay (RIPA) lysis buffer containing 1x protease inhibitor and 1x phosphatase inhibitor cocktail and identified by using a Tris-Tricine gel (each lane was loaded 40 μg protein) [11 (link)]. The Aβ protein levels were detected with 6E10 monoclonal antibody (1 : 500; BioLegend). Species-specific β-actin was the internal control detected with mouse β-actin monoclonal antibody (60008-1-Ig; 1 : 2000; Proteintech®). Anti-mouse IgG HRP-linked antibody was the secondary antibody (No. 7076, 1 : 3000; Cell Signaling Technology). Blots were visualized by standard enhanced chemiluminescence (ECL; NCI4106; Thermo Fisher). The protein signals were quantified with Gel-Pro Analyzer 4.

Anti-Aβ₄₂ antibody was purchased from BioLegend (San Diego, CA; USA, 6E10 monoclonal antibody for the detection of amyloid plaque burden in brain sections of APP/PS1 mice at 10 months of age, when most, if not all, of the human APP is cleaved into Aβ). Anti-Iba1 and anti-Nrf2 antibodies were purchased from GeneTex (Irvine, CA, USA). Anti-mouse/rabbit IgG HRP-conjugated antibody was purchased from Santa-Cruz Biotechnology (Santa Cruz, CA, USA). Anti-rabbit/mouse fluorescent-labeled antibody was purchased from Thermo Fisher Scientific (Wathham, MA, USA). The original rabbit anti-MetO antibody was produced in our lab as previously described [23 (link)].