Rabbit igg hrp

In this work we used the following primary antibodies: mouse monoclonal anti-STAT3 (1:1000) (BD Transduction Laboratories, 610189), mouse monoclonal anti-phosphoSTAT3 (1:100) (pY705) (BD Transduction Laboratories, 612356), rabbit polyclonal anti-PARP p85 Fragment pAb (1:500) (Promega, G7341), rabbit polyclonal anti-Mcl-1 (1:500) (Cell Signaling, 5453P) and goat polyclonal anti-caspase 3 (Santa Cruz Biotechnology Inc., sc-1225). To study autophagy we used the following primary antibodies: rabbit polyclonal anti-LC3 (1:1000) (Novus Biologicals, NB100-2220SS), mouse monoclonal anti-p62 (1:1000) (BD Transduction Laboratories, 610832) and rabbit polyclonal anti-Beclin1 (1:1000) (Cell Signaling, 3738S).
Monoclonal mouse anti-β-actin (1:10000) or anti-GAPDH (1:1000) (Santa Cruz Biotechnology Inc., sc-137179) were used as markers of equal loading” in “were used as markers of equal loading. Goat polyclonal anti-mouse IgG-horseradish peroxidase (HRP) (Santa Cruz Biotechnology Inc., sc-2005) rabbit polyclonal anti-goat IgG-HRP (Santa Cruz Biotechnology Inc., sc-2768) and rabbit IgG-HRP (Santa Cruz Biotechnology Inc., sc-2004) were used as secondary antibodies. All the primary and secondary antibodies used in this study were diluted in a PBS- 0.1% Tween 20 solution containing 3% BSA.

The fucoidan powder sample was a gift (Hi-Q oligo-fucoidan^®) from Hi-Q Marine Biotech International, Ltd. (Taiwan), and fucoidan of Fucus vesiculosus was purchased from Sigma-Aldrich Co. (St. Louis, MO). Fucoidan was dissolved in double-distilled H₂O (ddH₂O) and stirred at 25°C for 30 min. The dissolved solution was filtered using 0.22-μm sterile filters (Millipore®). The proteasome inhibitor MG-132 was purchased from CalBiochem, EMD Millipore (Billerica, MA). Recombinant human TGFβ1 was purchased from R&D Systems, Inc. (Minneapolis, MN). PD98059 was purchased from Cell Signaling (Beverly, MA). LY294002 was purchased from A.G. Scientific (San Diego, CA). Antibodies against p-Akt (S473), Akt, p-Smad2/3, Smad2/3, Smad4, β-actin, TGFβR I, and TGFβR II, normal rabbit IgG, rabbit IgG-HRP, mouse IgG-HRP, rat IgG-HRP (for immunoblotting) and protein A/G plus-agarose were obtained from Santa Cruz Biotechnology (Santa Cruz, CA); TGFRI, ubiquitin, Smurf2, Smad7, p-FAK (Y397), and rabbit IgG-HRP were purchased from GeneTex (Hsinchu, Taiwan, or USA); and TGFRII was from Cell Signaling (Beverly, MA).