Antibodies for AMPK and Tau Signaling

Antibodies directed against AMPKα1/α2, ACC, pSer⁷⁹-ACC were obtained from Cell Signaling Technology. Anti-AMPK α1 and anti-AMPK α2 antibodies were from R&D Systems. Anti p-Thr¹⁷²AMPK antibody and p-Thr¹⁷²AMPK blocking peptide were obtained from Santa Cruz Biotechnology24 (link). Anti-actin antibody was from BD Transduction Laboratory. Anti-MAP2, anti-α-tubulin and anti-acetylated-α-tubulin antibodies were from Sigma. Mouse monoclonal antibodies PHF1 (tau pSer³⁹⁶/Ser⁴⁰⁴)50 (link), CP13 (tau pSer²⁰²)51 (link), 2E12 or RZ3 (tau pThr²³¹)5 (link), DA9 (total tau aa102-140)36 (link), DA31 (total tau aa220-240)36 (link) and MC1 (conformation dependent antibody that recognizes only tau in a pathological conformation)52 (link) were previously described. 12E8 monoclonal antibody was obtained from Dr. Seubert (Elan Pharmaceuticals, San Francisco, CA; tau pSer^262/356)53 (link). As total tau antibody in the Western-blot experiments, we used tau Cter (homemade well characterized antibody recognizing the 11 amino-acids in the c-terminal part of tau27)54 (link). Tau5 (total tau) and AT180 (tau pThr231) were from Invitrogen.
AICAR, metformin, PD98059, H-89, LY294002 were purchased from Tocris; roscovitin and MARK/Par1 inhibitor from Merck; Compound C was from Santa-Cruz Biotechnology and rapamycin was from Cell Signaling Technology.