Cytotoxicity detection kit ldh assay

Cell viability was determined by using the cell proliferation reagent WST-1 (ROCHE, Penzberg, Germany) in a colorimetric assay. The WST-1 reagent was directly added to the media and was incubated for 4 h. After the reaction was complete, the plate was then immediately read at 450 nm. A reference reading was performed at 630 nm.
A cytotoxicity Detection Kit (LDH assay) (ROCHE, Penzberg, Germany) was used to measure the LDH released into the culture medium upon cell death due to damage to the plasma membrane. The culture media was directly collected from NSCs incubated with and without different concentrations of Aβ. The LDH content was assessed by an enzyme-linked immunosorbent assay (ELISA) and was read at an absorbance of 490 nm in a multimode microplate reader (BioTek Instruments, Winooski, VT, USA) with a reference wavelength of 630 nm.

Cell viability and death were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction (Sigma-Aldrich, St. Louis, MO, USA) and Cytotoxicity Detection Kit (LDH-assay)(Roche, Basel, Switzerland) assays, essentially as described in ref. 57 (link). The Anoikis resistance assay was performed with Corning Costar Ultra-Low attachment 24-well plates (Corning Life Science, Tewksbury, MA, USA). Briefly, 48 h post transfection, cells were plated and after another 24 h, cellular viability was assessed either manually by counting cells with loss of membrane integrity, or Trypan blue exclusion followed by automatic cell counting (Cedex XS Analyzer, Roche).