Rabbit polyclonal anti dr5 antibody

CFZ was purchased from Selleck Chemicals (Houston, TX) and was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 mM, and aliquots were stored at −80°C. Stock solutions were diluted to the desired final concentrations with growth medium just before use. CHX and actinoimycin D were purchased from Sigma Chemical Co. (St. Louis, MO). Human recombinant TRAIL was purchased from PeproTech, Inc. (Rocky Hill, NJ). Rabbit polyclonal anti-DR5 antibody was purchased from ProSci, Inc. (Poway, CA). Mouse monoclonal anti-DR4 antibody (B-N28) was purchased from Diaclone (Stamford, CT). Mouse monoclonal anti-caspase-3 was purchased from Imgenex (San Diego, CA). Rabbit anti-caspase-8 and anti-PARP antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA). Mouse monoclonal anti-FLIP antibody (NF6) was purchased from Alexis Biochemicals (San Diego, CA). Rabbit polyclonal Mcl-1 and Bcl-X_L and mouse monoclonal Bcl-2 antibody were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Rabbit polyclonal anti-β-actin antibody was purchased from Sigma Chemical Co.

Western blotting was carried out as previously described with a minor modification [39 ]. The antibodies used in the study were as followed: Rabbit polyclonal anti-DR5 antibody was purchased from ProSci, Inc (Poway, CA); mouse monoclonal anti-DR4 antibody (B-N28) was purchased from Diaclone (Stamford, CT); rabbit polyclonal anti-caspase-8, rabbit anti-poly (ADP-ribose) polymerase (PARP) and rabbit monoclonal anti-p-4EBP1 antibodies were purchased from Cell Signaling Technology, Inc (Beverly, MA); rabbit monoclonal anti-p-eIF4E (pS209) was purchased from Epitomics (USA); and rabbit polyclonal anti-GAPDH antibodies was purchased from Trevigen, Inc (Gaithersburg, MD). Quantification of signal intensity (IOD, integral optical density) was performed with Gel-Pro Analyzer software (Version 4.0). Expression change was indicated by IOD ratio of targeted protein before and after treatments. And the intensity was normalized by GAPDH signal. All detections were repeated for three independent times.

The MEK inhibitor U0126 was purchased from LC Laboratories (Woburn, MA). The JNK inhibitor SP600125 was purchased from Enzo Life Sciences, Inc./Biomol (Farmingdale, NY). Rabbit polyclonal anti-DR5 antibody was purchased from ProSci Inc. (Poway, CA). Mouse monoclonal anti-DR4 antibody (B-N28) was purchased from Diaclone (Stamford, CT). Protein-A/G plus-agarose, rabbit polyclonal anti-TRAF2 (sc-7187) and Fra-1 antibodies were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). Rabbit polyclonal antibody against HA tag was purchased from Abgent (San Diego, CA). Mouse monoclonal anti-MMP1 antibody was purchased from NeoMarkers, Inc. (Union City, CA). Antibodies against actin and Flag-tag and anti-Flag M2 affinity gel were purchased from Sigma-Aldrich (St. Louis, MO). Mouse monoclonal anti-TRAF2 (#558890) and anti-FADD (#556402) antibodies were purchased from BD Biosciences (San Jose, CA). Mouse monoclonal anti-caspase-8 (#9746), rabbit monoclonal anti-caspase-8 (#4790), rabbit polyclonal ant-FADD (#2782) and all other antibodies were all purchased from Cell Signaling Technology, Inc. (Beverly, MA). Anti-HA matrix agarose was purchased from Thermo Scientific/Pierce (Rockford, IL).