Abstract
We developed fixed-cell multi-well plate immunoassays that increase the throughput and ease of quantification for questions formerly assessed by immunoblot scanning. The assays make use of the now abundant antibodies designed to recognize receptor subtypes and posttranslationally modified signaling proteins. By optimizing permeabilization and fixation conditions, mainly based on specific cell types, the assay can be adapted to the study of many different antigens of importance to hormonal and neurotransmitter signaling scenarios.
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Acknowledgments
This work was jointly supported over the last 9 years by the National Institute of Environmental Health Sciences at the National Institutes of Health, the Department of Defense Breast Cancer Initiative, the Sealy Memorial Endowment Fund, the University of Texas Medical Branch Center for Addiction Research, the American Institute for Cancer Research, and the Passport Foundation, all in the USA.
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Watson, C.S. et al. (2014). Multi-well Plate Immunoassays for Measuring Signaling Protein Activations/Deactivations and Membrane vs. Intracellular Receptor Levels. In: Castoria, G., Auricchio, F. (eds) Steroid Receptors. Methods in Molecular Biology, vol 1204. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-1346-6_11
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DOI: https://doi.org/10.1007/978-1-4939-1346-6_11
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