Effective flow cytometric phenotyping of cells using minimal amounts of antibody

Deepak Sharma; Mark R. Eichelberg; Jill D. Haag; Amanda L. Meilahn; Matthew J. Muelbl; Kathy Schell; Bart M. G. Smits; Michael N. Gould

doi:10.2144/0000113854

BioTechniques (Jul 2012)

Effective flow cytometric phenotyping of cells using minimal amounts of antibody

Deepak Sharma,
Mark R. Eichelberg,
Jill D. Haag,
Amanda L. Meilahn,
Matthew J. Muelbl,
Kathy Schell,
Bart M. G. Smits,
Michael N. Gould

Affiliations

Deepak Sharma: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Mark R. Eichelberg: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Jill D. Haag: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Amanda L. Meilahn: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Matthew J. Muelbl: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Kathy Schell: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Bart M. G. Smits: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA
Michael N. Gould: 1Department of Oncology, University of Wisconsin School of Medicine and Public Health, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, 1400 University Avenue, Madison, WI, USA

DOI: https://doi.org/10.2144/0000113854
Journal volume & issue: Vol. 53, no. 1
pp. 57 – 60

Abstract

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Here we introduce a modified antibody staining method that uses up to 80% less antibody for flow cytometry. We demonstrate this method for the detection of antigens expressed at high, moderate, or low levels in mouse and rat lymphocytes as well as mouse mammary epithelial cells. We obtained reproducibly accurate results for the detection of up to seven parameters for activation induced-proliferation, cell cycle analysis, and phenotyping of cell-surface and intracellular antigens.

Published in BioTechniques

ISSN: 0736-6205 (Print); 1940-9818 (Online)
Publisher: Taylor & Francis Group
Country of publisher: United Kingdom
LCC subjects: Science: Biology (General)
Website: https://www.tandfonline.com/journals/ibtn20

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Abstract

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