Anti telomerase reverse transcriptase antibody

Expression of hTERT in immortalized POMECs was examined by western blotting. Cells were digested by trypsin and rinsed twice with PBS. RIPA buffer containing 1 mM phenylmethylsulfonyl fluoride (Solarbio, Beijing, China) was used for cellular protein extraction. Equivalent amount of proteins samples was separated by 10% SDS PAGE and transferred to a polyvinylidene fluoride membrane. After washing and blocking, membranes were incubated with anti-β-actin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or anti-telomerase reverse transcriptase antibody (1:1,000, Abcam) at 4°C overnight. Blots were detected with Western Light kit (Advansta, Menlo Park, USA) after membranes were incubated with secondary antibodies (horseradish-peroxidase-labeled goat anti-mouse or rabbit IgG [1:5,000, Bioss, Beijing, China)] for 1 h at RT.

Soybean phosphatidylcholine (S100-PC) was purchased from Lipoid GmbH (Ludwigshafen, Germany). DC-chol, 2-Iminothiolane hydrochloride (2-IT, Traut’s reagent), CTX and sulforhodamine B (SRB) were provided by Sigma-Aldrich (Saint Louis, MO, USA). Mal-PEG2000-DSPE was obtained from Avanti Polar Lipids (Alabaster, AL, USA). Anti-Transferrin Receptor antibody (OX-26) and anti-Telomerase reverse transcriptase antibody were purchased from Abcam (Cambridge, UK). Sephadex G-50 and Sepharose CL-4B were provided by Science and Technology Development Co., Ltd. (Beijing, China). Centriprep-10 concentrators were purchased from Millipore Amicon (Bedford, MA, USA). NanoOrange® Protein Quantitation Kit was purchased from Invitrogen (California, USA). YOYO-1 and LysoTracker Red were purchased from Molecular Probes (Eugene, OR, USA). pIRES2-EGFP (pEGFP) and pIRES2-EGFP/hTERTC27 (pC27) were purified using a E.Z.N.A.TM Fastfilter Plasmid Maxi Kit (Omega, Norcross, GA, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies (Tokyo, Japan). Lipofectamine™ 2000 Reagent (Lipo2000) was purchased from Invitrogen (California, USA). pGL3 Luciferase Reporter Vectors (pGL3-luc) and Luciferase Reporter Assay Kit were purchased from Promega (Madison, WI, USA).