Abstract
Measurement of protein expression in live, intact cells using flow cytometry (FC) has been employed for several decades in the areas of immunology, cell biology, and molecular biology. More recently, this technique has found appreciation in applied scientific fields, including cancer biology and endocrinology, to serve as a tool for identifying cells more likely to respond to specific treatments. FC, also referred to as fluorescence-activated cell sorting (FACS), is an antibody-based method that provides the user with an ability to identify proteins expressed on surfaces of cells as well as in the cytoplasm, including steroid hormone receptors. This technique is most useful for examining specific cell types in a heterogeneous population and therefore can be used to identify cells more likely to respond to treatments based on expression of the appropriate receptor. Isolation of purified subpopulations for further manipulation and investigation of functional capacity is also possible using a cell sorter, which uses similar technology to isolate cells for use by the researcher. This is especially important for studying responses of less abundant cell populations in tissues that express high levels of a target protein or receptor of interest. Furthermore, FACS analysis is clinically useful to identify and isolate responsive cell populations, which may be less appreciable in whole tissues because of the diluting effects of surrounding, nonresponding cell types. Immune cells are commonly utilized as a source of cell populations in the FC technique and have previously been shown to express steroid hormone receptors and respond to steroid hormone treatment. Here, we demonstrate that FC is a useful tool for identifying immune cells expressing steroid hormone receptor protein. This method can also be easily expanded to include other, nonimmune cell populations to address specific research questions related to steroid hormone receptor biology.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Herzenberg LA, Sweet RG, Herzenberg LA. Fluorescence-activated cell sorting. Sci Am 1976;234(3):108–17.
Herzenberg LA, Parks D, Sahaf B, Perez O, Roederer M, Herzenberg LA. The history and future of the fluorescence activated cell sorter and flow cytometry: a view from Stanford. Clin Chem 2002;48(10):1819–27.
McCoy JP, Jr. Basic principles of flow cytometry. Hematol Oncol Clin North Am 2002;16(2):229–43.
McCoy JP, Jr., Carey JL, Krause JR. Quality control in flow cytometry for diagnostic pathology. I. Cell surface phenotyping and general laboratory procedures. Am J Clin Pathol 1990;93(4 Suppl 1):S27–37.
McCoy JP, Jr., Overton WR. A survey of current practices in clinical flow cytometry. Am J Clin Pathol 1996;106(1):82–6.
Berki T, Kumanovics G, Kumanovics A, Falus A, Ujhelyi E, Nemeth P. Production and flow cytometric application of a monoclonal anti-glucocorticoid receptor antibody. J Immunol Methods 1998;214(1–2):19–27.
Marchetti D, Van NT, Gametchu B, et al. Flow cytometric analysis of glucocorticoid receptor using monoclonal antibody and fluoresceinated ligand probes. Cancer Res 1989;49(4):863–9.
Butts CL, Shukair SA, Duncan KM, et al. Progesterone inhibits mature rat dendritic cells in a receptor-mediated fashion. Int Immunol 2007;19(3):287–96.
Butts CL, Shukair SA, Duncan KM, Harris CW, Belyavskaya E, Sternberg EM. Effects of dexamethasone on rat dendritic cell function. Horm Metab Res 2007;39(6):404–12.
Butts CL, Shukair SA, Duncan KM, Harris CW, Belyavskaya E, Sternberg EM. Evaluation of steroid hormone receptor protein expression in intact cells using flow cytometry. Nucl Recept Signal 2007;5:e007.
Boldizsar F, Palinkas L, Czompoly T, Bartis D, Nemeth P, Berki T. Low glucocorticoid receptor (GR), high Dig2 and low Bcl-2 expression in double positive thymocytes of BALB/c mice indicates their endogenous glucocorticoid hormone exposure. Immuno-biology 2006;211(10):785–96.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Butts, C.L., Sternberg, E.M. (2009). Flow Cytometry as a Tool for Measurement of Steroid Hormone Receptor Protein Expression in Leukocytes. In: McEwan, I.J. (eds) The Nuclear Receptor Superfamily. Methods in Molecular Biology™, vol 505. Humana Press. https://doi.org/10.1007/978-1-60327-575-0_3
Download citation
DOI: https://doi.org/10.1007/978-1-60327-575-0_3
Publisher Name: Humana Press
Print ISBN: 978-1-60327-574-3
Online ISBN: 978-1-60327-575-0
eBook Packages: Springer Protocols