Secondary antibodies conjugated with horseradish peroxidase

The cell lysates were prepared as described previously [20 (link)]. Briefly, the Tris-based buffer containing phosphatase inhibitors and protease inhibitors (Roche, Basel, Switzerland) was applied and cleared with centrifugation. The protein in soluble fractions was quantified using a BCA kit (Thermo) and standard western blotting was performed using PVDF membrane (Merk Millipore, Boston, MA, USA). The antibodies used were as follows: α-tubulin (sc-23948) and PARP1/2 (sc-7150) were purchased from Santa Cruz (Santa Cruz, CA, USA). SCD1 (ab19862) was purchased from Abcam (Cambridge, UK). Cleaved active caspase-3 (#9661) was purchased from Cell Signaling Technology (Danvers, MA, USA). The secondary antibodies conjugated with horseradish peroxidase were purchased from Cell Signaling Technology. The signals were visualized using SuperSignal^® Femto (Thermo) and X-ray films (Agfa, Mortsel, Belgium).

Mouse monoclonal antibodies for SNAP-25 (SP-12, diluted 1:2000) and syntaxin 1 (HPC-1, 1:2000) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit monoclonal anti-VAMP 2 antibodies (D6O1A, 1:1000) were purchased from Cell Signaling Technology (Danvers, MA, USA). Secondary antibodies conjugated with horse radish peroxidase (1:5000) were purchased from Cell Signaling Technology.