Rabbit anti ulk

For Western blotting, equivalent amounts of sample protein were separated by 10% SDS-PAGE and transferred to nitrocellulose membrane (Macherey Nagel, Düren, Germany). After blocking with 5% BSA in TBS-T (20 mM Tris pH 7.6, 150 mM NaCl and 0.02% Tween-20), the membrane was immunoblotted with primary antibodies overnight at 4 °C. The following primary antibodies were used: rabbit anti-LRRC8A (1 μg/mL, [4 (link)] kindly supplied by T. J. Jentsch); rabbit anti-p-ULK (1:1000, Cell Signaling, Frankfurt am Main, Germany; #14202); rabbit anti-ULK (1:1000, cell signaling, #8054); rabbit anti-p-Akt^Ser473 (1:1000, Cell Signaling; #4060); rabbit anti-Akt^pan (1:1000, Cell Signaling; #4685); and rabbit anti-GAPDH (1:2500, Cell Signaling; #2118) antibodies. The corresponding peroxidase-labeled secondary antibody (1:10,000, Jackson ImmunoResearch, Ely, UK) was used. Signals were detected using an enhanced chemiluminescence reagent (HRP juice; PJK GmbH, Kleinblittersdorf, Germany) and a ChemiSmart5000 digital imaging system (Vilber-Lourmat, Collégien, France). Densitometrical quantification was performed with the Fiji software [64 (link)].