Ratiometric and nonratiometric Ca2+ indicators for the assessment of intracellular free Ca2+ in a breast cancer cell line using a fluorescence microplate reader

Robinson, J. A., Jenkins, N. S., Holman, N. A., Roberts-Thomson, S. J. and Monteith, G. R. (2004) Ratiometric and nonratiometric Ca2+ indicators for the assessment of intracellular free Ca2+ in a breast cancer cell line using a fluorescence microplate reader. Journal of Biochemical And Biophysical Methods, 58 3: 227-237. doi:10.1016/j.jbbm.2003.11.002


Author Robinson, J. A.
Jenkins, N. S.
Holman, N. A.
Roberts-Thomson, S. J.
Monteith, G. R.
Title Ratiometric and nonratiometric Ca2+ indicators for the assessment of intracellular free Ca2+ in a breast cancer cell line using a fluorescence microplate reader
Journal name Journal of Biochemical And Biophysical Methods   Check publisher's open access policy
ISSN 0165-022X
Publication date 2004-01-01
Year available 2004
Sub-type Article (original research)
DOI 10.1016/j.jbbm.2003.11.002
Open Access Status Not yet assessed
Volume 58
Issue 3
Start page 227
End page 237
Total pages 11
Editor C. F. Chignell
S. Hjerten
Place of publication Netherlands
Publisher Elsevier BV
Language eng
Subject C1
270199 Biochemistry and Cell Biology not elsewhere classified
780105 Biological sciences
Abstract Transporters of Ca2+ are potential drug targets and Ca2+ is a useful signal in the assessment of G-protein-coupled receptor activation. Assays involving the assessment of intracellular Ca2+ using microplate readers most often use Ca2+ indicators which do not exhibit a spectra shift on Ca2+ binding (e.g. fluo-3). Indicators that do exhibit a spectral shift upon Ca2+ binding (e.g. fura-2) offer potential advantages for the calibration of intracellular Ca2+ levels. However, experimental limitations may limit the use of ratiometric dyes in microplate readers capable of screening. In this study, we compared the assessment of intracellular Ca2+ in adherent breast cancer cells using ratiometric and nonratiometric Ca2+ indicators. Our results demonstrate that both fluo-3 and fura-2 detect ATP dose-dependent increases in intracellular Ca2+ in the MCF-7 breast cancer cell line and that some of the limitations in the use of fura-2 appear to be overcome by the use of glass bottom microplates. The calibrated intracellular Ca2+ levels derived using fura-2 are consistent with those from microscopy and cuvette-based studies. Fura-2 may be useful in microplate studies, where cell lines with different properties are compared or where screening treatments lead to differences in the number of cells or dye loading. (C) 2003 Elsevier B.V. All rights reserved.
Keyword Biochemistry & Molecular Biology
Biophysics
Calcium
Fura-2
Mcf-7
Screening
Membrane Calcium-pump
Plasma-membrane
Entry
Homeostasis
Modulation
Diversity
Sparks
Biochemical Research Methods
Q-Index Code C1
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2005 Higher Education Research Data Collection
School of Pharmacy Publications
 
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