Optimized Ki-67 staining in murine cells: a tool to determine cell proliferation
| Autor: | Lars Eichhorn, P Wurst, Elmar Endl, C Graefe, Andreas Hoeft, Jan Lukas Kleiner, I Panetas, Christina Weisheit, Z Abdulla, Annkristin Heine, Stilla Frede, Christian Kurts |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Flow cytometry Mice 03 medical and health sciences 0302 clinical medicine Cell Line Tumor Tumor Cells Cultured Genetics medicine Animals Humans Molecular Biology Cell Proliferation Fixation (histology) Cell Nucleus Staining and Labeling biology medicine.diagnostic_test Chemistry Cell growth General Medicine Cell cycle Flow Cytometry Staining Cell biology Mice Inbred C57BL Ki-67 Antigen 030104 developmental biology Microscopy Fluorescence Cell culture 030220 oncology & carcinogenesis Ki-67 biology.protein Antibody Cell Division |
| Zdroj: | Molecular Biology Reports. 46:4631-4643 |
| ISSN: | 1573-4978 0301-4851 |
| DOI: | 10.1007/s11033-019-04851-2 |
| Popis: | The reliable analysis of the cell cycle status has become increasingly relevant for scientific and clinical work, especially for the determination of tumor cell growth. One established method to characterize the proliferation activity of cells is the analysis of the Ki-67 protein. Ki-67 is expressed in the nucleus during the whole cell cycle except for the G0 phase. Several different protocols exist for the examination of the Ki-67 protein in tissue and cell culture, but most of them are defined for human cells. For the analysis of the Ki-67 protein in murine tissue and cell culture there is a variety of protocols existing which recommend different fixation and permeabilization reagents or special kits. In this study, we established a reliable protocol for Ki-67 staining in murine cells and tissue based on PFA fixation, which can be used not only for flow cytometry but also for immunofluorescence microscopy analysis. We tested our protocol successfully with three different Ki-67 anti-mouse antibodies in cell culture, regenerating liver tissue and mouse melanoma tumor to demonstrate the general applicability. |
| Databáze: | OpenAIRE |
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