Goat anti mouse rabbit igg

Treated cells were lysed using the Mammalian Cell Lysis kit (Sigma-Aldrich). Equal amounts of protein were electrophoresed in a sodium dodecyl sulfate-polyacrylamide gel under reducing conditions followed by transfer to PVDF membranes (Millipore, IPVH00010). The blots were blocked with 5 % nonfat dry milk in PBS (137 mM NaCl; 2.7 mM KCl; 10 mM Na2HPO4; 2 mM KH2PO). The western blots were then probed with respective antibodies. The protein amounts loaded were normalized according to the β-actin signal using Mouse Anti-β-Actin antibody (Sigma-Aldrich). The secondary antibodies were HRP conjugated to goat anti-mouse/rabbit IgG (Santa Cruz, sc-2005 and sc-2004).

Treated cells were lysed using the Mammalian Cell Lysis kit (Sigma, MCL1-1KT). Equal amounts of the proteins were electrophoresed in a sodium dodecyl sulfate-polyacrylamide gel (12%) under reducing conditions followed by transfer to PVDF membranes (Millipore, IPVH00010). The blots were blocked with 5% nonfat dry milk in phosphate-buffered saline (PBS; 137 mM NaCl; 2.7 mM KCl; 10 mM Na₂HPO₄; 2 mM KH₂PO₄). The western blots were then probed with antibodies recognizing the indicated proteins. The protein amounts loaded were normalized according to the ACTB/β-actin signal, using an anti-ACTB antibody (Sigma, A5441). The secondary antibodies were HRP conjugated to goat anti-mouse/rabbit IgG (Santa Cruz, sc-2005 and sc-2004).