Anti aldh1 mouse monoclonal antibody clone 44 aldh

Western blotting was performed as described previously [50 (link)]. Briefly, 1 × 10⁵ of ALDH^high cells and ALDH^low cells derived from HEC-1 cells, siRNA transfected HEC-1 cells and HSF1 plasmid transfected HEC-1 cells were lysed in 100 μl of SDS sample buffer. Anti-ALDH1 mouse monoclonal antibody (clone: 44/ALDH, BD Pharmingen) was used at 1000-times dilution. Anti-HSP27 rabbit polyclonal antibody (Abcam, Cambridge, UK), anti-HSF1 rabbit polyclonal antibody (Sigma-Aldrich, Saint Louis, MO), anti-HSF1 pSer320 rabbit polyclonal antibody (Abcam, Cambridge, UK), anti-HSF1 pSer326 rabbit polyclonal antibody (Abcam, Cambridge, UK), anti-HSF1 pSer230 rabbit polyclonal antibody (Santa cruz, Dallas, TX), anti-HSF1 pSer303 rabbit polyclonal antibody (Sigma-Aldrich, Saint Louis, MO) and anti-HSF1 pSer307 rabbit polyclonal antibody (Santa cruz, Dallas, TX) were used at 1000-times dilution. Anti-β-Actin mouse monoclonal antibody (Sigma-Aldrich, Saint Louis, MO) was used at 2000-times dilution. Anti-mouse IgG+IgM and anti-rabbit IgG and IgM second antibodies (KPL) were uses at 2000-times dilution. The membrane was visualized with Chemiluminescent HRP Substrate (Milipore Corporation, Bilerica, MA) according to the manufacturer's protocol, and pictures were taken by an Odyssey^® Fc Imaging System (LI-COR, Lincoln, NE).

Western blots were performed as described previously [25 (link)]. Briefly, 1 × 10⁵ of ALDH1^high cells and ALDH1^low cells were lysed in 40 μL of SDS sample buffer. Anti-ALDH1 mouse monoclonal antibody (clone: 44/ALDH; BD Biosciences, San Jose, CA, USA) was used at 500-times dilution. Anti-β-actin and anti-α-tubulin mouse monoclonal antibodies (Sigma-Aldrich) were used as loading controls at 2000-times dilution. Anti-mouse IgG and anti-rabbit IgG second antibodies (KPL, Gaithersburg, MD, USA) were used at 2000-times dilution. The membrane was visualized with Chemiluminescent HRP Substrate (Millipore, Billerica, MA, USA) according to the manufacturer's protocol, and pictures were taken by an Odyssey Fc imaging System (LI-COR, Lincoln, NE, USA).