Rat bdnf elisa kit

The medium after treatment with NGF peptides was collected at 24 h of incubation and diluted with the provided sample buffer (1:1) immediately prior to the assay. Levels of BDNF in PC12 cells supernatants were measured with Rat BDNF ELISA kit (ERBDNF, Thermofisher, Waltham, MA, USA).

Levels of BDNF were determined using Thermo Scientific’s Rat BDNF ELISA kit per the manufacturer’s instructions. Briefly, the sample (100 µl) was added to its appropriate well, and incubated for 2.5 h at room temperature. The solution was discarded, and the wells were washed 4 times with 1× Wash Buffer. Next, 100 µl of 1× prepared biotinylated antibody was added to each well and incubated for 1 h at room temperature with gentle shaking. 100 µl of prepared Streptavidin-HRP solution was then added to each well and incubated for 45 min at room temperature with gentle shaking. The solution was than discarded and the wells were washed 4 times with 1× Wash Buffer. Finally, 100 µl of TMB Substrate was added to each well and incubated for 30 min at room temperature in the dark with gentle shaking. Each plate was evaluated within 30 min of stopping the reaction. Optical density OD values were measured at the wavelength of 450 nm, and the OD values of supernatant and serum in hippocampal homogenate were measured using a multi-function enzyme labelling analyzer.

Levels of Aβ42 in conditioned medium of H4‐APPswe were determined with the Human/Rat beta‐Amyloid (42) ELISA Kit, High Sensitivity from Wako Chemicals USA, Inc. Medium‐containing human recombinant Aβ42, collected at 6 and 20 h incubation, was diluted with the provided sample buffer immediately prior to the assay. Levels of BDNF in neuronal culture supernatants were measured with Rat BDNF ELISA kit Thermofisher.