Goat anti rabbit horse radish peroxidase conjugate

Celastrol with purity greater than 98% was purchased from Alexis Biochemicals (San Diego, CA, United States). RPMI 1640, 0.4% trypan blue vital stain, and antibiotic-antimycotic mixture were obtained from Invitrogen (Carlsbad, CA, United States). Propidium iodide (PI), MTT, Tris, glycine, NaCl, SDS, BSA, and β-actin antibody (mouse monoclonal) was obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, United States). FBS was purchased from BioWest (Miami, FL, United States). Bortezomib (Velcade, PS341) was purchased from LC Laboratories (Woburn, MA, United States). Nuclear extraction and DNA binding kits was obtained from Active Motif (Carlsbad, CA, United States). Caspase-Glo 3/7 Assay kit was purchased from Promega (Madison, WI, United States). Rabbit polyclonal antibodies against Poly(ADP-ribose) polymerase (PARP), MMP-9, caspase-3, and goat anti-rabbit-horse radish peroxidase (HRP) conjugate and goat anti-mouse HRP conjugate were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Rabbit polyclonal antibody against CXCR4 was purchased from Abcam (Cambridge, MA, United States). CXCL12 was purchased from ProSpec-Tany TechnoGene Ltd. (Rehovot, Israel). ELISA kits for mouse IL-6 and TNF-α were purchased from R&D Systems Inc. (Minneapolis, MN, United States).

γ-tocotrienol with purity more than 97% was obtained from Davos Life Science, Singapore in the form of Naturale³ γ-tocotrienol. γ-tocotrienol was dissolved in dimethylsulfoxide (DMSO) as a 10 mM stock solution and stored at 4°C for the in vitro experiments. Further dilution was done in cell culture medium as required. Human recombinant vascular endothelial growth factor (VEGF) and growth factor reduced (GFR) matrigel matrix were purchased from BD Biosciences (CA, USA). Mayer's hematoxylin solution, eosin Y Solution, MTT, Tris, glycine, NaCl, SDS, BSA, and β-actin antibody were purchased from Sigma-Aldrich (St. Louis, MO). Medium 200, low serum growth supplement (LSGS), DMEM medium, fetal bovine serum (FBS), 0.4% trypan blue vital stain, and antibiotic-antimycotic mixture were obtained from Invitrogen (Carlsbad, CA). Rabbit polyclonal antibodies to phospho-AKT (Ser 473), phospho-mTOR (Ser 2448), phospho-p70 S6 Kinase (Thr 389), phospho-p70 S6 Kinase (Thr421/Ser424), AKT, mTOR, p70S6K1 and CD31 were purchased from Cell Signaling Technology (Beverly, MA). Antibodies against VEGF, Ki-67, PARP, cleaved caspase-3, goat anti-rabbit-horse radish peroxidase (HRP) conjugate, and goat anti-mouse HRP were purchased from Santa Cruz Biotechnology (San Diego, CA).