Rabbit anti alpha beta tubulin

Cell lysates were prepared and resolved as described^{12 (link)}. Primary antibodies used were: rabbit anti-PPP1R15A (10449-1-AP, 1:1000; Proteintech, Manchester, UK), rabbit anti-phospho-eIF2α (3597, 1:1000; Cell Signaling), mouse anti-actin (ab3280, 1:1000; Abcam), rabbit anti-ATF4 (C-20, 1:500; SantaCruz, Santa Cruz, CA, USA), mouse anti-total eIF2α (AHO0802, 1:1000; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA); guinea pig anti-insulin (ab7842, 1:1000; Abcam), rabbit anti-GAPDH (14C10) (2118, 1:1000; Cell Signaling), rabbit anti-UCP1 (ab10983, 1:1000; Abcam), rabbit anti-CHOP (1:1000; kind gift from Professor David Ron, Cambridge, UK), rabbit anti-alpha/beta tubulin (2148, 1:1000; Cell Signalling, Hitchin, UK).

Western blots were performed as described [11 (link)]. For fluorescent westerns, cell lysates were prepared with phosphatase inhibitors (β-Glycerophosphate and Sodium orthovanadate) and a protease inhibitor (cOmplete™, Mini Protease Inhibitor Cocktail, Roche 04693124001), membranes were read using a Li-Core Odyssey laser scanner, and results analysed using Image Studio Lite software.
The following primary antibodies were used: mouse anti-total ERK (p44/42 MAPK (Erk1/2) L34 F12 Cell Signalling 4696s), mouse anti-Kras (CPTC-KRAS4B-2, DSHB) rabbit anti-dpERK, (p44/42 MAPK, Cell Signalling 4370), rabbit anti-alpha/beta tubulin (Cell Signalling 2148), goat anti-rabbit IgG-HRP (Jackson), goat anti-mouse IgG-HRP (Jackson). Secondary antibodies for fluorescent westerns: donkey anti-mouse 800 (Rockland 610–732-124), goat anti-rabbit 680 (Molecular Probes A21076).