Anti stim1 pab

Hippocampal regions from WT mice at 3.5 month were dissected, homogenized, and solubilized at 4 °C for 1 h in lysis buffer (1 % CHAPS, 137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4, 1.4 mM KH2PO4, pH 7.2, 5 mM EDTA, 5 mM EGTA, 1 mM PMSF, 50 mM NaF, 1 mM Na3VO4 and protease inhibitors). Primary hippocampal neurons were collected, homogenized and solubilized in the same buffer on DIV14. The total protein lysates were separated by SDS-PAGE and analyzed by Western blotting with anti-STIM1 pAb (1:1000, Cell Signaling, 4916 s), anti-STIM2 pAb (1:1000, AnaSpec, 54681), anti-Phospho-CaMKII (1:2000, Abcam, ab171095), anti-CaMKII (1:1000, Chemicon, MAB8699), anti-PSD95 (1:1000, Cell Signaling, 3450 s), anti-actin clone C4 (1:1000, Millipore, MAB1501) and anti-tubulin (1:1000, DSHB, E7-c). HRP-conjugated anti-rabbit (1:2000, DAKO, P0448) and anti-mouse (1:2000, DAKO, P0447) secondary antibodies. Analysis of the data was performed using Quantity One from BioRad software. The mean density of each band was normalized to actin or tubulin signal in the same sample and averaged. All Western blots replicated 3–7 times.