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
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.
