A rapid microwell fluorescence immunoassay for cellular protein detection
- 22 September 2008
- journal article
- Published by Springer Science and Business Media LLC
- Vol. 10, 83-89
- https://doi.org/10.1251/bpo146
Abstract
In this paper, we describe a simple, rapid, specific, sensitive, and reliable method, the FICP method (Fluorescence Immunoassay for Cellular Protein detection) which is readily applicable to the detection of proteins directly on cells cultured in 96-well plates. In order to illustrate this method, we report on the detection of two different proteins, the cell cycle proteins cyclin D1 and p21CIP1/WAF1, in untreated and 2-cyclopenten-1-one treated breast cancer cells. When the FICP method was compared with Western blot procedure, FICP was found to be superior for many characteristics. By using this method, we were able to quantify biological effects of a specific compound on protein levels in non-lysed cells and perform statistical analysis. Therefore, we believe this screening assay could be very useful for detecting poorly expressed proteins and for drug development.This publication has 23 references indexed in Scilit:
- Specific subcellular localization of siRNAs delivered by lipoplex in MCF-7 breast cancer cellsBiochimie, 2007
- Transfected cell microarrays: an efficient tool for high-throughput functional analysisExpert Review of Proteomics, 2007
- 15-Deoxy-Δ12,14-prostaglandin J2 inhibits G2-M phase progression in human breast cancer cells via the down-regulation of cyclin B1 and survivin expressionBreast Cancer Research and Treatment, 2006
- Cell microarrays and RNA interference chip away at gene functionNature Genetics, 2005
- Limitations of current proteomics technologiesJournal of Chromatography A, 2005
- Regulation of Gene Expression by a Metabolic EnzymeScience, 2004
- Culture of Human Normal Brain and Malignant Brain Tumors for Cellular, Molecular, and Pharmacological StudiesPublished by Springer Science and Business Media LLC ,2003
- Cell-biological applications of transfected-cell microarrays.Trends in Cell Biology, 2002
- Prostaglandin A2 Specifically Represses Insulin-Like Growth Factor-I Gene Expression in C6 Rat Glioma Cells*Endocrinology, 1997
- New Colorimetric Cytotoxicity Assay for Anticancer-Drug ScreeningJNCI Journal of the National Cancer Institute, 1990