β-Amyloid (1-42), rat

Cell experiment:

After treating the hippocampal slices with 20 μM β-Amyloid (1-42) for 4 h, supernatant is replaced by fresh H-ASCF/3 (0.9 mL/chamber) to which 0.1 mL MTT stock solution (5 mg/mL H-ACSF/3) is added (MTT final concentration: 0.5 mg/mL). The chamber is left to rest for 15 min without carboxygenation. To stop further reduction of MTT, the medium (H-ACSF/3) is removed. The slices are transferred into 96-well plate, then pure DMSO (100 μL/slice/well) is added for dissolving formazane from the slices. (30 min in a 96-well plate). Then 70 μL DMSO solution from each slice (well) is transferred into another 96-well plate. The optical density (OD) of the dissolved formazane is measured at 550 and 620 nm[1].

References:

[1]. Mozes E, et al. A novel method for the rapid determination of beta-amyloid toxicity on acute hippocampal slices using MTT and LDH assays. Brain Res Bull. 2012 Apr 10;87(6):521-5.
[2]. Lagunes T, et al. Abeta(1-42) induces abnormal alternative splicing of tau exons 2/3 in NGF-induced PC12 cells. An Acad Bras Cienc. 2014 Dec;86(4):1927-34.