Journal: Nature protocols
Article Title: A 3D human neural cell culture system for modeling Alzheimer’s disease
Figure Lengend Snippet: Analysis of soluble Aβ levels in the enriched FAD ReN cells by Western Blot and Aβ ELISA a. Western Blot of Aβ levels in the conditioned medium collected from undifferentiated enriched ReN-G and -GA cells. The conditioned medium was collected from T25 flask after culturing the cell for 2 days. 6E10 anti-Aβ detected ~4 kDa Aβ monomer bands in the media collected from the enriched FAD ReN cells (ReN-GA10, ReN-GA2). The 6E10 antibody also detected sAPPα (~100 kDa), which would represent the amount of full-length APPs in these cell lines. b . Increased Aβ40 and 42 levels in undifferentiated FAD ReN-GA2 cells, detected through Aβ ELISA (***, p
Article Snippet: Greiner Bio-One 96-well PS uClear black TC plate, flat bottom (Greiner Bio-One, 5665-5090) MatTek glass bottomed 96-well plates (MatTek, cat. no. P96GC-0-5-F) Corning® Black/Clear bottom 96-well plates (Corning, cat. no. 3340) Lab-Tek II 8-well chambered cover glass slides (Thermo Scientific, cat. no. 155409) Lab-Tek 16-well glass chamber slides (Thermo Scientific, cat. no. 178599) MatTek glass bottomed 35 mm plates (MatTek, cat. no. P35G-0-7-C or P35G-1.5-10-C) T25 cell culture flasks (BD Biosciences, cat. no. 353082) 6-well cell culture plates (Fisher Scientific/Corning life sciences, cat. no. 353046) Cell culture inserts, 0.4 μm pore size (Fisher Scientific/Falcon, cat. no. 353095) 24-well Falcon companion plates for cell culture inserts (Fisher Scientific/Falcon, cat. no. 353504) Cell culture inserts, 0.4 μm pore size (Greiner Bio-One, cat. no. 662654) ▲ CRITICAL Both cell culture inserts (Falcon and Greiner Bio-One) work well for thick-layer culture for biochemical analysis.
Techniques: Western Blot, Enzyme-linked Immunosorbent Assay