Amyloid beta 42 human elisa kit

Aβ ELISA was performed as previously described [11 (link)]. In brief, mouse forebrains were homogenized in TBST and centrifuged. The supernatant was collected for TBST soluble Aβ, and the pellet was further homogenized in GuHCl for TBST insoluble Aβ. Aβ levels in the mouse forebrain and culture medium were quantified by Amyloid beta 40 Human ELISA Kit (Invitrogen, KHB3482) and Amyloid beta 42 Human ELISA Kit (Invitrogen, KHB3544). TNF-α, IL-1β, and IL-6 levels in mouse forebrain and culture medium were quantified by Mouse TNF alpha ELISA Kit (ab208348, abcam), Mouse IL-1 beta ELISA Kit (ab197742, abcam) and Mouse IL-6 ELISA Kit (ab222503, abcam).

Weekly, 1.4e7 BE(2)-m17 cells per line were plated in a well of a six-well plate without antibiotics (hygromycin and puromycin) and with daily media changes. On the fourth day, supernatant was harvested and supplemented to a final concentration of 1.0 mM of an irreversible serine protease inhibitor, AEBSF (Thermo Fisher Scientific), then clarified at 10,000 g for 10 min at room temperature. Resulting supernatant was transferred to a new tube and stored at -80°C until analysis. Cells in the plate were lysed for or RNA extraction using the Quick-RNA miniprep kit (Zymo Research, Irvine, CA, USA), which were stored at -80°C until analysis.
For the first two sets of APP metabolite experiments, after 3 weeks of samples had been stored, supernatants were diluted fourfold and assayed with the Amyloid beta 40 Human ELISA Kit and either the Amyloid beta 42 Human ELISA Kit or the Amyloid beta 42 Human ELISA Kit Ultrasensitive (Invitrogen), according to the manufacturer's instructions. For the second two sets, supernatants were diluted fourfold and assayed with the V-PLEX Plus Abeta Peptide Panel 1 (6E10) Kit and the sAPPalpha/sAPPbeta Kit (Meso Scale Discovery), according to the manufacturer's protocol.

The effects of saponins on the expression levels of AD-related genes, p-tau and Aβ(1–42), were measured using specific ELISA kits (Amyloid beta 42 Human ELISA Kit Invitrogen and Tau (Total) Human ELISA Kit) following the manufacturer’s instructions. SHY-5Y cells were cultured under the stated conditions in presence and absence (control) of various nontoxic concentrations of saponins. After treatment and incubation with saponins, lysate was prepared and processed in 96-well microplates for the experiments following the kit’s instructions. Absorbance was measured at 450 nm using an ELISA microplate reader. Data were analyzed and compared according to the manufacturer’s instructions.

Hippocampal tissue (10–15 mg) was homogeneized in eight volumes (v/w) cold TBS containing protease inhibitors, briefly sonicated and cleared by centrifugation at 15,000 × g at 4°C for 45 min. The levels of Aβ42 peptides were assessed in duplicate using the Amyloid beta 42 Human ELISA Kit (Invitrogen, Carlsbad, USA). Aβ42 concentrations were extrapolated from a standard curve of synthetic human Aβ42 (range: 15.6–1,000 pg/ml) and corrected for protein concentration, as determined by the Lowry assay, and are expressed as pg Aβ42 per mg protein.