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Quantitative Flow Cytometry-Based Assays for Measuring Constitutive Secretion

  • David E. Gordon
  • Amber S. Shun-Shion
  • Asral W. Asnawi
  • Andrew A. PedenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2233)

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.

Key words

Constitutive secretion Flow cytometry VSV-G GPI hGH Transport Assay Trafficking Biosynthetic transport 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • David E. Gordon
    • 1
  • Amber S. Shun-Shion
    • 2
  • Asral W. Asnawi
    • 2
    • 3
  • Andrew A. Peden
    • 2
    Email author
  1. 1.Department of Cellular and Molecular PharmacologyUniversity of California San FranciscoSan FranciscoUSA
  2. 2.Department of Biomedical Science, Centre for Membrane Interactions and DynamicsUniversity of SheffieldSheffieldUK
  3. 3.Faculty of Medicine and Health SciencesUniversiti Sains Islam MalaysiaNegeri SembilanMalaysia

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