Rabbit monoclonal anti ptak1

Total cellular protein was prepared using RIPA buffer containing a protease inhibitor cocktail and protein concentration was measured using the BCA assay as described [4 (link)]. In brief, protein was analyzed by SDS-PAGE, transferred to nitrocellulose (Schleicher and Schuell, Keene, NH). After blocking, the blots were incubated overnight at 4 °C with the appropriate primary antibody in Tris-buffered saline with 1 % Tween (TBST) and 5 % BSA. Secondary antibodies were applied for 1 h at room temperature. Specific proteins were detected using the enhanced chemiluminescence system (Amersham, Arlington Heights, IL). The primary antibodies that were used were rabbit monoclonal anti-pSmad 1/5, rabbit monoclonal anti-pSmad2, rabbit monoclonal anti-Smad2, rabbit monoclonal anti-pTAK1, rabbit monoclonal XIAP, rabbit anti-monoclonal p-p65, rabbit monoclonal anti-activated caspase-3, rabbit polyclonal anti-BMPRII, and rabbit monoclonal anti-EGR-1 (Cell signaling Technology, Danvers MA), Rabbit monoclonal anti-TAK1 (Invitrogen, Grand Island NY), rabbit anti-actin, an affinity isolated antigen specific antibody (Sigma, Saint Louis, MO), rabbit monoclonal anti-Id1 and rabbit monoclonal anti-Id3 (Calbioreagents, San Mateo, CA), rabbit polyclonal anti-GAPDH (Sigma, St. Louis, MO) and rabbit polyclonal pMEK-1/2 (Cell Signaling, Danvers MA).