Anti tata binding protein antibody

Mouse monoclonal antibodies against tubulin and actin were from Sigma and anti-phospho-ERK from Cell Signaling Technology. Polyclonal antibodies for the EMT markers, Bcl-2 family members, PARP, cleaved caspase 3, ERK, phospho-ERK, AKT, phospho-AKT (Serine 473), were from Cell Signaling Technology. Anti-TATA binding protein (TBP) antibody was from Abcam. Antibodies for Myc and EPOR (C20) were from Santa Cruz Biotechnology. For immunohistochemistry, the EPOR antibody GM1201 was used (Aldevron GmbH).

Isolation of cytoplasmic and nuclear proteins has been described previously [28 (link)]. Protein extracts were electrophoresed through SDS polyacrylamide gels. Gels were electrophoretically transferred to Immobilon PVDF membranes (Millipore, Bedford, MA) and reacted with the indicated antibodies. Immunoreactive proteins were visualized by the chemiluminescence detection system ImmunoStar LD (Wako, Osaka, Japan). The following antibodies were used: anti-RCAN1 and anti-beta-actin antibodies (Sigma-Aldrich, St. Louis, MO), anti-phospho-c-JUN (Ser63), and anti-c-JUN antibodies (Cell Signaling Technology), and anti-TATA binding protein (TBP) antibody (Abcam, Cambridge, UK).

CEP (HPLC purity: 99.1%, batch number: 0483686) was purchased from the Cayman Chemical Company. Methylprednisolone (MP) and verapamil (VP) were obtained from Sigma-Aldrich (St. Louis. Mo., USA). CEP, MP, and VP were dissolved in ethanol and stored at 4℃ until use. ConA was purchased from Seikagaku Kogyo Co. Tokyo, Japan. The WST-8 reagent was provided by Dojindo Molecular Technologies, Inc., Tokyo, Japan. The BD Cytometric Bead Array Human Th1/Th2/Th17 Cytokine Kit, FITC mouse anti-human CD4, APC mouse anti-human CD8 and CD25, and Alexa Fluor^® 488 mouse anti-human Foxp3 antibodies were obtained from BD Biosciences (San Jose, CA, USA). The anti-P-glycoprotein antibody was purchased from Kamiya Biomedical Company, Seattle, USA (1 μg/mL, # MC-208). Anti-GR antibody was obtained from Santa Cruz Biotechnology, INC, Texas, USA (G-5, dilution:1:1000, # sc-393232). The anti-β-actin antibody was provided by Proteintech Group, Rosemont, USA (1:5000, # 66009-1-lg). The anti-TATA binding protein (TBP) antibody was purchased from Abcam (Cambridge, UK; dilution:1:1000, # ab818). RPMI-1640 and fetal bovine serum (FBS) were obtained from Gibco BRL (Grand Island, NY, USA). All other reagents used were of the highest quality available from commercial vendors.