Irdye 680rd goat anti rat igg

Whole cell lysates were prepared using NP-40 lysis buffer (50 mM Tris-HCL pH 7.5, 150 mM NaCl, 1% NP-40 (v/v), 1 mM EDTA pH 8) supplemented with cOmplete protease inhibitor and PhosSTOP phosphatase inhibitor cocktails (Roche, Basel, Switzerland). A total of 30 μg of protein per sample was resolved on NuPAGE 4-12% Bis-Tris pre-cast polyacrylamide gels (Novex, Thermo Fisher Scientific), transferred onto iBLOT2 Polyvinylidene Difluoride (PVDF) membranes (Invitrogen, Thermo Fisher Scientific) and blocked using Odyssey blocking buffer (LiCor, Lincoln, NE, USA). Primary antibodies used in western blot analysis were BIM_S/L/EL (ALX-804-527, Enzo Life Sciences, New York, NY, USA), MCL-1 (5453S, Cell Signalling Technologies, Danvers, MA, USA), BCL-X_L (2764, Cell Signalling Technologies), Cleaved Caspase 3 (9661, Cell Signalling Technologies), and β-tubulin (ab6046, Abcam, Cambridge, UK). Secondary antibodies used were IRDye®680RD Goat anti-rat IgG (H + L)(926-68076, LiCor) and IRDye®800CW Goat anti-rabbit IgG (H + L)(926-32211, LiCor). Full western blot images can be found in the Supplementary File.

For immunocytochemistry the following antibodies were used: Alexa Fluor® 488 Donkey anti-Goat, Alexa Fluor® 488 Donkey anti-Mouse, Alexa Fluor® 488 Donkey anti-Rabbit, Alexa Fluor® 488 Donkey anti-Rat, Alexa Fluor® 555 Donkey anti-Mouse, Alexa Fluor® 555 Donkey anti-Rabbit, Alexa Fluor® 555 Donkey anti-Rat, Alexa Fluor® 647 Donkey anti-Mouse, Alexa Fluor® 647 Donkey anti-Rabbit (Invitrogen).
For immunoblotting, the following antibodies were used: IRDye® 680RD Donkey anti-Mouse IgG, IRDye® 680RD Goat anti-Rat IgG, IRDye® 800CW Donkey anti-Rabbit IgG (LI-COR). For visualization of the the Dig(T)40 probe, FITC-labeled anti-Digoxigenin-AP, Fab fragments from sheep (Roche) was used.