Eriksson, Staffan
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
Abstract
Thymidine kinase 1 (TK1) is a DNA salvage enzyme involved in the synthesis of thymidine triphosphate needed during S phase. Although TK1 has been utilized as a cell proliferation marker for many years no well-characterized antibodies are available. The preparation and properties of two types of poly- and monoclonal anti-TK1 peptide antibodies are described and they are used to determine the levels of TK1 in intact cells. Expression of TK1, c-fos, cyclin B1, Ki67, phosphorylated histone H3, phosphorylated ribosomal protein S6, as well as bromodeoxyuridine (BrdU) incorporation in human normal dermal fibroblast cultures were studied with high-content ArrayScan (TM) fluorescence microscopy. The levels of TK1 increased 6-7h after serum re-addition to starved cells as they passed through G1, S and G2/M phases, which was earlier than the increase in Ki67 protein levels and before BrdU incorporation was detected. Thus, a population of activated G1 cells with high TK1 and low Ki67 expression could be identified and their role in cell proliferation can now be clarified. (C) 2009 Elsevier GmbH. All rights reserved.
Keywords
Cell cycle; Biomarkers; Thymidine kinase; G1 cells; Monoclonal antibodies; Immunofluorescence
Published in
European Journal of Cell Biology
2009, volume: 88, number: 12, pages: 779-785
Publisher: ELSEVIER GMBH, URBAN & FISCHER VERLAG
SLU Authors
UKÄ Subject classification
Animal and Dairy Science
Veterinary Science
Publication identifier
- DOI: https://doi.org/10.1016/j.ejcb.2009.06.005
Permanent link to this page (URI)
https://res.slu.se/id/publ/49005