Trap silver staining telomerase detection kit

pSIECs (passage 3), ZYM-SIEC02 cells (passage 50) and A549 cells (a lung adenocarcinoma cell line) were trypsinized, washed twice with PBS, and pelleted by centrifugation at 2,000 rpm for 5 min. Telomerase activity was analyzed using a TRAP-silver staining Telomerase Detection Kit (KeyGEN Biotech, Nanjing, China) according to the manufacturer’s instructions.

DMEM/F12 and FBS were purchased from Gibco. EGF, ITS-G and Lipofectamine Plus were products of Invitrogen. The WST-1 Cell Proliferation and Cytotoxicity Assay Kit was obtained from Beyotime, Shanghai, China. The Annexin V-FITC Apoptosis Detection Kit was a product of Calbiochem, Darmstadt, Germany. The TRAP-silver staining Telomerase Detection Kit was purchased from KeyGEN Biotech, Nanjing, China. The following primary antibodies were used: Pan-cytokeratin (clone AE1/AE3, AbD Serotec, Oxford, UK), mouse anti-Cytokeratin 18 (clone CY90, Sigma, St. Louis, MO, USA), rabbit anti-Sucrase-isomaltase (clone H-123, Santa Cruz, Heidelberg, Germany), rabbit anti-E-cadherin antibody (Genscript), mouse anti-OCLN (Clones 1G, AbD, Oxford, UK), rabbit anti-villin and anti-ZO-1 (Bioss, Beijing, China).
Healthy unsuckled 1-day-old Landrace piglets were purchased from the pig farm of Northwest Agriculture & Forestry University.
Nude mice at 4 weeks old of age were purchased from the Experimental Animal Center of The Fourth Military Medical University (Xi'an, China) and maintained in pathogen-free conditions.