Synaptophysin antibody

Human Aβ_1–42 was purchased from EMD Millipore Corporation (Billerica, MA, USA). Ori was obtained from Chengdu Must Bio-Technology Company (Sichuan, China). Neurobasal media and B27 were bought from Life Technologies Corporation. MAP-2 antibody was purchased from Abcam (Cambridge, MA, USA). COX-2 antibody was bought from Santa Cruz Biotechnology (Santa Cruz, USA). PSD-95 antibody, synaptophysin antibody and CREB antibody were obtained from CST (Cell Signaling Technology, USA). 4', 6'-diamidino-2-phenylindole (DAPI), VDAC1, BDNF, p-TrkB, TrkB, p-CREB and β-actin antibodies were purchased from Bioworld Technology (Bioworld, USA). Lamin B1 antibody and HRP-conjugated secondary antibodies were also obtained from Bioworld.

The quantitative Western blot was utilized in the present studies. We used antibody AT8 (55 ​kDa, 1:1000; Invitrogen, Carlsbad, CA) to detect the amounts of Tau phosphorylated at serine 202 and threonine 205 (Tau-PS202/PT205) amino acid. IL-6 antibody (24 ​kDa, 1:1000; Cat. # ab6672, Abcam, Cambridge, MA) was used to recognize IL-6. Postsynaptic density (PSD)-95 antibody (95 ​kDa, 1:1000; Cell Signaling, Danvers, MA), synaptophysin antibody (38 ​kDa, 1:1000; Cell Signaling) and N-cadherin antibody (140 ​kDa, 1:1000; Cell Signaling) were used to detect the protein amounts of PSD-95, synaptophysin, and N-cadherin, respectively. A β-Actin antibody (42 ​kDa, 1:5000; Sigma) was used to detect β-Actin. The quantification of Western blot was accomplished as described in other studies (Dong et al., 2009 (link)). In brief, we analyzed the signal intensity via Quantity One image analysis program (Bio-Rad, Hercules, CA). Two steps were used to quantify the Western blots. At the first step, β-Actin was used to standardize protein amounts (e.g., calculating the ratio of PSD-95 as compared to β-Actin amount), limiting the differences in the protein amount loaded. At the second step, we expressed the protein amounts obtained from the treatment as a percentage to the control condition.