Multiparametric Flow Cytometry of Murine Hematopoietic Cells

Single cell suspensions obtained from colony-forming assays or hematopoietic organs from mice were surface stained with monoclonal antibodies: CD34 PE-Cy7(581), CD38 APC (HIT2), CD10 PE/APC (HI10a), CD45RA PerCP-Cy5.5 (HI100), CD90 APC-Cy7 (5E10), CD117 PE-Cy7 (104D2), CD71 APC (CY1G4), CD33 PE (WM53), CD14 APC (M5E2), CD115 BV421 (4D2-1E4), CD15 PeCy5 (W6D3), CD66b PerCP-Cy5.5 (G10F5), CD19 PE-Cy7 (HIB19), IgM APC-Cy7 (MHM-88), CD45 Biotin (HI30), CD45 PE-Cy7/V500 (30-F11) (Biolegend), and CD235a BV421 (HIR2) (BD Biosciences). Streptavidin PerCP-Cy5.5/V450 (Biolegend) was used as a secondary antibody. BD LSRFortessa and FlowJo were used for flow cytometry and analyses, respectively. The gating strategy was published earlier.^{31 (link)}

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Bohler S., Afreen S., Fernandez-Orth J., Demmerath E.M., Molnar C., Wu Y., Weiss J.M., Mittapalli V.R., Konstantinidis L., Schmal H., Kunze M, & Erlacher M. (2020). Inhibition of the anti-apoptotic protein MCL-1 severely suppresses human hematopoiesis. Haematologica, 106(12), 3136-3148.

Publication 2020

CD34 PE-Cy7 CD38 APC CD45RA PerCP-Cy5.5 CD90 APC-Cy7 CD117 PE-Cy7 CD71 APC CD33 PE CD14 APC CD115 BV421 CD66b PerCP-Cy5.5 CD19 PE-Cy7 IgM APC-Cy7 CD45 Biotin ( CD235a BV421 (HIR2)

Antibody Assays Biotin Cd235a Cd66b Cd71 Cd90 Cell Cy5 5 Flow cytometry Hematopoietic Mice Monoclonal antibodies Streptavidin

Corresponding Organization :

Other organizations : University of Freiburg, University Medical Center Freiburg, German Cancer Research Center, Heidelberg University

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Variable analysis

independent variables

Monoclonal antibodies used for surface staining: CD34 PE-Cy7, CD38 APC, CD10 PE/APC, CD45RA PerCP-Cy5.5, CD90 APC-Cy7, CD117 PE-Cy7, CD71 APC, CD33 PE, CD14 APC, CD115 BV421, CD15 PeCy5, CD66b PerCP-Cy5.5, CD19 PE-Cy7, IgM APC-Cy7, CD45 Biotin, CD45 PE-Cy7/V500, CD235a BV421

dependent variables

Cell surface marker expression levels measured by flow cytometry

control variables

Single cell suspensions obtained from colony-forming assays or hematopoietic organs from mice
Streptavidin PerCP-Cy5.5/V450 used as a secondary antibody

positive controls

Not specified

negative controls

Not specified

Annotations

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