Abstract
Isolation of cells expressing transfected cDNAcan be extremely difficult and tedious if the efficiency of expression is low (less than 1–5%). If the receptor being selected for is a cell surface or integral membrane protein at least partly exposed to the exterior, then it is possible to use the extremely powerful technique of fluorescence-activated cell sorting (FACS) to select these cells from a receptor-negative background.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sklar, L. A. and Finney, D. A. (1982) Analysis of ligand-receptor interactions with the fluorescence activated cell sorter. Cytometry 3, 161–165.
Machines, D. G., Green, D. K., Harmer, A., Nairn, E. G., and Fink, G. (1983) Neuroendocrine receptor—ligand binding using quantitative video—intensification microscopy and fluorescence—activated cell sorting. Quart. J. Exp. Physiol. 68, 463–474.
Due, C, Linnet, K., Langeland Johansen, N., and Olsson, L. (1985) Analysis of insulin receptors on heterogeneous eukaryotic cell populations with fluorochrome—conjugated insulin and fluorescence—activated cell sorter. Diabetologia 28, 749–755.
Chatelier, R. C., Ashcroft, R. C., Lloyd, C. J., Nice, E. C., Whitehead, R. H., Sawyer, W. H., and Burgess, A. W. (1986) Binding of fluoresceinated epidermal growth factor to A431 cell sub-populations studied using a model—independent analysis of flow cytometric fluorescence data. EMBO J. 5, 1181–1186.
Tubiana, N., Mishal, Z., Le Caer, F., Seigneurin, J. M., Berthiox, Y., Martin, P. M., and Carcassonne, Y. (1986) Quantification of oestradiol binding at the surface of human lymphocytes by flow cytometry. Br. J. Cancer 54, 501–504.
Newman, R., Domingo, D., Trotter, J., and Trowbridge, I. (1986) Selection and properties of a mouse L-cell transformant expressing human transferrin receptor. Nature 304, 643–645.
Ellis, L., Clauser, E., Morgan, D. O., Edery, M., Roth, R. A., and Rutter, W. J. (1986) Replacement of insulin receptor tyrosine residues (1162 and 1163) by site-directed mutagenesis compromises insulin—stimulated kinase activity and uptake of 2-De-oxyglucose. Cell 45, 721–732.
Clark, S., Cheng, D. J., Hsuan, J. J., Haley, J. D., and Waterfield, M. D. (1988) Loss of three major auto phosphorylation sites in the EGF—receptor does not block the mitogenic action of EGF. J. Cell Physiol. 133, 421–425.
Waterfield, M. D., Mayes, E. L. V., Stroobant, P., Bennett, P. L. P., Young, S., Goodfellow, P. N., Banting, G. S., and Ozanne, B. (1982) A monoclonal antibody to the human epidermal growth factor receptor. J. Cell Biochem. 20, 149–161.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1990 The Humana Press Inc.
About this protocol
Cite this protocol
Clark, S. (1990). Fluorescence-Activated Cell Sorting According to Receptor Density. In: Walker, J.M., Pollard, J.W., Walker, J.M. (eds) Animal Cell Culture. Methods in Molecular Biology, vol 5. Humana Press. https://doi.org/10.1385/0-89603-150-0:559
Download citation
DOI: https://doi.org/10.1385/0-89603-150-0:559
Publisher Name: Humana Press
Print ISBN: 978-0-89603-150-0
Online ISBN: 978-1-59259-492-4
eBook Packages: Springer Protocols