Peroxidase labeled goat anti rabbit and goat anti mouse

For Western blotting, total protein extracts were obtained in cell lysis buffer containing 150 mM NaCl, 1 mM EDTA, 1% NP-40, 50 mM Tris (pH 8.0) as well as phosphatase and protease inhibitors. Protein concentrations were determined by BCA staining (#23225, Thermo Fisher Scientific, Hennigsdorf, Germany) as compared to BSA concentration standard. Following SDS polyacrylamide gel electrophoresis, proteins were blotted on nitrocellulose membranes, as described previously [55 (link)].
Several primary antibodies were derived from Cell Signaling Technology (Danvers, MA, USA): caspase-3 (9662, rabbit, 1:1000), caspase-8 (9746, mouse, 1:1000), caspase-9 (9502, rabbit, 1:1000), Mcl-1 (4572, rabbit, 1:1000), Bcl-w (2724, rabbit, 1:1000) and Bcl-2 (2872, rabbit, 1:1000). Other primary antibodies were derived from Santa Cruz Biotech (Dallas, TX, USA): Bcl-x_L (sc-8392, mouse, 1:1000) and β-actin (sc-47778, mouse, 1:1000). As secondary antibodies, peroxidase-labeled goat anti-rabbit and goat anti-mouse were used (Dako, Hamburg, Germany; 1:5000).

For Western blotting, total protein extracts were obtained by cell lysis in 150 mM NaCl, 1 mM EDTA, 2 mM PMSF, 1 mM leupeptin, 1 mM pepstatin, 0.5% SDS, 0.5% NP-40 and 10 mM Tris-HCl, pH 7.5. Western blotting on nitrocellulose membranes was performed as described previously.^{45 (link)} Primary antibodies were: Cleaved caspase-3 (9664, rabbit, 1 : 1000, Cell Signaling, Danvers, MA, USA), caspase-3 proform (9662, rabbit, 1 : 1000, Cell Signaling), Mcl-1 (sc-12756, mouse, 1 : 200, Santa Cruz Biotech, Dallas, TX, USA), Bcl-2 (sc-492, rabbit; 1 : 200, Santa Cruz Biotech), Bax (sc-20067, mouse, 1 : 200, Santa Cruz Biotech), GAPDH (sc-32233, mouse, 1 : 1000, Santa Cruz Biotech), Puma (ab33906, rabbit, 1 : 1000, Abcam, Cambridge, UK), XIAP (no. 2042, rabbit, 1 : 1000, Cell Signaling), p53 (sc-126, rabbit, 1 : 500, Santa Cruz Biotech), ERK (no. 4695, rabbit, 1 : 1,000, Cell Signaling), pERK (no. 9101, rabbit, 1 : 1,000, Cell Signaling), and Bim (no. 559685, rabbit, 1 : 200, BD Biosciences, Heidelberg, Gemany). Secondary antibodies were: peroxidase-labeled goat anti-rabbit and goat anti-mouse (Dako, Hamburg, Germany; 1 : 5000).

For Western blotting, total proteins were extracted by lysing the cells in cell lysis buffer, containing 150 mM NaCl, 1 mM EDTA, 1% NP-40, 50 mM Tris-HCl, pH 8.0, and inhibitors for proteases and phosphatases. After SDS polyacrylamide gel electrophoresis, proteins were transferred by electro blotting to nitrocellulose membranes.
The following primary antibodies were used: Cleaved caspase-3 (9664, rabbit, 1:1000; Cell Signaling, Danvers, MA, USA); Caspase-8 (9746, mouse, 1:1000; Cell Signaling), Caspase-9 (9502, rabbit, 1:1000; Cell Signaling); p21 (sc-6246, mouse, 1:200; Santa Cruz Biotech, Dallas, TX, USA); p19 (sc-1063, rabbit, 1:200; Santa Cruz Biotech); β-actin (sc-47778, mouse, 1:200; Santa Cruz Biotech). The following secondary antibodies were used: peroxidase-labeled goat anti-rabbit and goat anti-mouse (Dako, Hamburg, Germany; 1:5000).