Abstract
Constitutive secretion is predominantly measured by collecting the media from cells and performing plate-based assays. This approach is particularly sensitive to changes in cell number, and a significant amount of effort has to be spent to overcome this. We have developed a panel of quantitative flow cytometry-based assays and reporter cell lines that can be used to measure constitutive secretion. These assays are insensitive to changes in cell number making them very robust and well suited to functional genomic and chemical screens. Here, we outline the key steps involved in generating and using these assays for studying constitutive secretion.
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Acknowledgments
We would like to thank the MRC (G120/952) and BBSRC (BB/L002841/1 and BB/S009566/1) for supporting this work. A. S. Shun-Shion was funded by the BBSRC White Rose DTP studentship (BB/J014443/1).
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Gordon, D.E., Shun-Shion, A.S., Asnawi, A.W., Peden, A.A. (2021). Quantitative Flow Cytometry-Based Assays for Measuring Constitutive Secretion. In: Niedergang, F., Vitale, N., Gasman, S. (eds) Exocytosis and Endocytosis. Methods in Molecular Biology, vol 2233. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1044-2_8
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DOI: https://doi.org/10.1007/978-1-0716-1044-2_8
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