Mouse monoclonal against β actin

Dithiothreitol, iodoacetamide, paraformaldehyde (PFA), CPT, proteases and phosphatases inhibitors cocktails were from Sigma-Aldrich. CHAPS was from Applichem (Darmstadt, Germany). DDP was from Pfizer (New York, NY, USA). Rabbit polyclonal antibody against SLIRP (Abcam Cambridge, UK), mouse monoclonal antibody against FLAG peptide (Sigma-Aldrich), mouse monoclonal against β-actin (Sigma-Aldrich), mouse monoclonal antibody against bcl-2 (Santa Cruz Biotechnology, Inc, Dallas, TX, USA) and rabbit polyclonal antibody against bcl-2 (Santa Cruz Biotechnology) were used. Lipofectamine2000 and BCA reagents were purchased from Invitrogen. Enhanced chemiluminescence (ECL) reagents from Pierce Biotechnology (Rockford, IL, USA), Protein A and Protein G agarose beads from Amersham Biosciences Europe (Milan, Italy), sequencing-grade modified trypsin from Promega (Madison, WI, USA) were used. Acetonitrile (ACN), formic acid (FA) and water for mass spectrometry were from Sigma. Water used in this study was deionized using a Milli-Q purification system (Millipore, Billerica, MA, USA).

HCT-116 WT and HCT-116 p21^−/− cells were maintained in McCoy’s 5A media and supplemented with 10% fetal bovine serum, gentamicin 50 µg/mL, and ciprofloxacin hydrochloride 5 µg/mL. Adenovirus antibodies used in this study consisted of mouse monoclonal antibody against E1A (M73) (Santa Cruz Biotech, Santa Cruz, CA, USA), E2A 72k DNA binding protein (B6-8) (provided as a kind gift by Dr. Arnold Levine, (Rutgers Cancer Institute of New Jersy), and rabbit polyclonal to Adenovirus type 5 capsid proteins (ab6982, Abcam, Cambridge, MA, USA). Cellular specific antibodies included mouse monoclonal against β-Actin (Sigma, St. Loius, MO, USA), mouse monoclonal anti-p21 (BD Biosciences, San Jose, CA, USA), mouse monoclonal anti-PCNA (Santa Cruz Biotech, Santa Cruz, CA, USA), chicken polyclonal anti-p21, and mouse monoclonal anti-BrdU (MoBU) (Invitrogen, Grand Island, NY, USA).