Journal
CLINICAL BIOCHEMISTRY
Volume 36, Issue 8, Pages 621-628Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0009-9120(03)00111-5
Keywords
telomerase activity; TRAP assay; silver staining; quantification; endometrium; hyperplasia; carcinoma
Categories
Ask authors/readers for more resources
Objectives : To develop a sensitive telomeric repeat amplification protocol (TRAP)-silver staining assay for telomerase activity quantification. Design and Methods : TRAP assays were performed by using a TRAPeze(R) telomerase kit with or without [alpha-P-32] -dCTP. Amplification products were electrophoresed in polyacrylamide, gels and detected by autoradiography or a modified silver staining protocol. Telomerase activity was quantified from radioactive counts or optical density of telomerase products from test extracts and controls. Results : TRAP-silver staining assay was at least as sensitive as radioactive TRAP assay and quantified telomerase activity within linearity from 10 to 3,000 cell equivalents. Both methods quantified a weak telomerase activity in normal endometrial glandular epithelial cells (GEC) and a strong increase in immortalized GEC. In human pathologic endometria (n = 24), telomerase activity was correlated with lesion seriousness and distinguished simple hyperplasias from nonhyperplasic or cancerous lesions. Conclusions : TRAP-silver staining assay is suitable for cell and tissue telomerase activity routine quantification. (C) 2003 The Canadian Society of Clinical Chemists. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available