Determination of Telomerase Activity in Tumors and Cancer Cell Lines - Divya, Kurunchi C.;Parthiban, M.
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The present study was undertaken to determine the presence of telomerase activity in normal and cancerous cell lines and tumor tissues by combined PCR-ELISA assay. Cell lines, canine mammary tumor tissues and human oral cancer tissues were processed for protein extraction. The concentration of the protein was determined by Bradford s method and used for Telomere Repeat Amplification Protocol (TRAP). The amplified products were electrophoresed on 20% non-denaturing polyacrylamide gel along with 10bp ladder. The presence of 6 base repeats starting from 50 nucleotides is considered as positive.…mehr

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Produktbeschreibung
The present study was undertaken to determine the presence of telomerase activity in normal and cancerous cell lines and tumor tissues by combined PCR-ELISA assay. Cell lines, canine mammary tumor tissues and human oral cancer tissues were processed for protein extraction. The concentration of the protein was determined by Bradford s method and used for Telomere Repeat Amplification Protocol (TRAP). The amplified products were electrophoresed on 20% non-denaturing polyacrylamide gel along with 10bp ladder. The presence of 6 base repeats starting from 50 nucleotides is considered as positive. Then the TRAP products were immobilized on to streptavidin coated microtitre plates via biotin - streptavidin interaction and then detected by anti-DNP antibody conjugated with horse radish peroxidase. The absorbance was measured at 450 nm and at 690 nm. Samples was regarded as telomerase positive if A is greater than 0.2, where A: net increase of absorbance for the sample = A sample A heat-treated sample.Telomerase serves as an efficient biomarker for diagnosis of tumor and cell lines serves as a model system to investigate the efficacy of telomerase targeted therapeutics for cancer.
Autorenporträt
Ms Kurunchi C Divya is presently working as Assistant Professor at Chettinad University, INDIA. Her area of interests are Molecular virology, cloning and expression studies and Animal cell culture.