Cells were lysed in RIPA buffer (20 mM Tris pH 7.5, 2 mM EDTA, 150 mM NaCl, 1 % NP40, 1 % SDS, 0.1 % TritonX100, 0.25 % Na-Deoxychalate, plus protease inhibitor) and the samples were incubated at 95 °C for 5 min. Protein lysates were separated on 10 % SDS-PAGE gels and transferred onto polyvinylidene difluoride (PVDF) membranes. As primary antibodies we used anti-Mx (mouse, M143 [31 (link)]), anti-RIG-I (mouse, AdipoGen), and anti-beta-actin (mouse, AC-15, Sigma-Aldrich). Primary antibodies were detected with peroxidase-conjugated secondary antibodies (GE Healthcare).
Free full text: Click here
