Ab193156

Cells were lysed with Cell Lysis Buffer (9803S, Cell Signaling Technology) containing 1 mM PMSF. Whole tissue lysates were generated using bead mills with 100 µl lysis buffer per 50 mg of tissue. SDS–PAGE was employed to separate proteins which were then blotted on a nitrocellulose membrane. Membranes were blocked in 5% skim milk dissolve in TBS with 0.1% Tween 20. Membranes were incubated with primary antibodies diluted in 5% skim milk in TBST overnight at 4°C. Following primary antibodies were used: anti‐β‐Actin (A5441, Sigma‐Aldrich, 1:3,000), anti‐BDH1 (15417‐1‐AP, Proteintech for Fig 1B, 1:2,000 or ab193156, Abcam, 1:1,000 for all other Western blots), anti‐SCOT (12175‐1‐AP, Proteintech, 1:1,000), anti‐β‐Tubulin (2146, Cell Signaling Technology, 1:2,500), anti‐VCP (ab11433, Abcam, 1:2,000). HRP‐conjugated secondary antibodies (DAKO) were used and chemiluminescence was detected by Aceglow ECL Western Blotting Substrate (VWR International).