Goat anti mouse sc2055

Western blots were performed essentially as described in [32 (link)] with the following modifications: a protease inhibitor cocktail (Roche, 1:25) was used and no additional dithiothreitol was added to the sample mix including loading dye. Aliquots of cell lysates were further applied for Western blot analysis as published previously [15 (link)]. The following proteins were detected by specific antibodies: OPN (M66105M, Meridian Life Science, Memphis, TN, USA), PI3Kinase p110 β (#3011), pPDK1 (#3438), pAkt (#13038P), pmTOR (#5536) (Cell signaling technology, Frankfurt, Germany), and β-actin (sc1615, Santa Cruz, Heidelberg, Germany), which served as internal loading control. The respective horseradish-peroxidase-conjugated secondary antibodies were donkey anti-goat (sc2020), goat anti-mouse (sc2055) (Santa Cruz), and goat anti-rabbit (#7074, Cell signaling technology).

Immunoblotting was done as described in (Yosifov et al. 2007 (link)), but a proteinase inhibitor cocktail (Roche, 25x) was used and additional dithiothreitol was not included to the sample mix and loading dye. Aliquots of the cell lysates were further analyzed as described before (Kovacheva et al. 2014 (link)). Specific antibodies detected the following proteins: integrin β3 (sc-46655) and β-actin (sc1615) (Santa Cruz, Heidelberg, Germany). Beta-actin served as internal loading control. The respective horseradish-peroxidase-conjugated secondary antibodies were goat anti-mouse (sc 2055) and donkey anti-goat (sc2020) (Santa Cruz, Heidelberg, Germany).