Ab73843

RP1-C was diluted to 40 μg/mL with coating buffer (15 mM Na₂CO₃, 35 mM NaHCO₃ [pH 9.6]), and the diluted rP1-C was used to coat an ELISA plate (100 μL per well) at 4°C overnight. The plate was washed three times with phosphate-buffered saline with Tween 20 (PBST) and blocked with 5% skim milk in PBST at 37°C for 2 h. After washing, 50 μL of vimentin (0.2 mg/mL; ab73843; Abcam, UK) was added to each well and incubated at 37°C for 2 h. Anti-vimentin antibody (1:1,000; ab223871; Abcam) and secondary antibody (1:10,000; BS12478; Bioworld, China) were subsequently incubated in sequence, and the A₄₅₀ was determined (Tecan; Infinite F50).

Primary mouse monoclonal antibodies were: anti-phospho-tau/MAP2 (12E8, provided by Dr Peter Seubert, Prothena Biosciences); GAPDH (G8795, Sigma); tubulin (T6199, Sigma); MAP2 (M1460, Sigma); vimentin (ab20346, Abcam); neurofilament (ab72995, Abcam); α-internexin (mab5224, Chemicon); fibrillarin (ab4566, Abcam). Primary rabbit monoclonal antibodies were anti-tau (ab76128, ab64193, Abcam). Primary rabbit polyclonal antibodies were: anti-tau (Dako, Abcam); FUS (11570, Proteintech); α-internexin (ab5354, Chemicon); ADF/cofilin (D8815, Sigma); vimentin (PAB9580, Abnova); EIF3e (ab36766, Abcam). Secondary antibodies were anti-mouse or anti-rabbit HRP-conjugated (Jackson ImmunoResearch); Alexa fluor-conjugated 488, 555 and 647 goat anti-mouse and anti-rabbit (Invitrogen). Recombinant proteins were: EIF3e (ab114769, Abcam); α-internexin (ab160346, Abcam); neurofilament (H00004747, Abnova); vimentin (ab73843, Abcam).