Mouse monoclonal antibody for beta actin

Excised C4-2 tumor tissue was homogenized as per the manufacturer’s protocol, using the Precellys™ tissue homogenizer system (Bertin Technologies, Montigny le Bretonneux, France). Proteins were extracted using a RIPA buffer as previously described [21 (link)]. Briefly, tumor tissue (100 mg) was homogenized in RIPA buffer with 1× protease inhibitor at a 1:4 (tissue: buffer) ratio using Precellys™ Tissue Homogenizing CKMix (Cat. # 3961-1-009) at 6000 rpm for two cycles of 20 s each with a 15 s break. Protein assay and quantification were performed as described previously [21 (link)]. Thirty to forty micrograms of protein were loaded per lane into 12% SDS-acrylamide gels. Mouse monoclonal antibody for beta actin was used as loading control (1:5000; Sigma-Aldrich, Oakville, ON, Canada) and rabbit polyclonal antibodies for VDR (1:200; Santa Cruz Biotechnology Inc., Dallas, TX, USA) were used to develop the immunoblots. Conjugated secondary antibodies (anti-mouse IRDye 800 at a dilution of 1:5000 and anti-rabbit IRDye 680 at a dilution of 1:20,000) were used and obtained from Cedarlane Laboratories (Burlington, ON, Canada).

N-acetyl cysteine (NAC) and mouse monoclonal antibody for beta-actin were purchased from Sigma (Atlanta, GA). Rabbit monoclonal antibody for phoshpo-p53 (Ser15), phospho-p38^MAPK (D3F9), cleaved PARP were purchased from Cell Signaling Technology (Tokyo, Japan). Rabbit polyclonal antibody for SESN3(bs6100R) was purchased from Bioss (MA, USA). Water soluble tetrazolium (WST) reagents was purchased from Takara Bio Inc. (Kyoto, Japan). Docetaxel were purchased from Wako (Kyoto, Japan). Cabazitaxel were purchased from Toronto Research Chemicals Inc (Ontario, Canada)