Multiparametric Flow Cytometry Analysis of Hematopoietic Cell Subsets

We first incubated cells with PE-conjugated anti-mouse antibodies directed against CD11b (clone M1/70), CD19 (clone 6D5), CD90.2 (clone 53-2.1), CD11c (clone N418), CD4 (clone GK1.5), CD8a (clone 53-6.7), CD127 (clone A7R34), CD49b (clone DX5), Ly-6G (clone 1A8), Ly-6C (clone HK1.4), TER-119 (clone TER-119) (all BioLegend). Then cells were stained with antibodies directed against c-kit (BioLegend, clone 2B8), sca-1 (BioLegend, clone D7), CD34 (BD Bioscience, clone RAM34), CD16/32 (BioLegend, clone 93), CD115 (eBioscience, clone AFS98). The term LSK refers to hematopoietic stem and progenitor cells based on their characteristic surface expression pattern (Lineage ^neg, Sca-1⁺, c-Kit⁺) [42 (link)], specified as Lin (CD11b, CD19, CD90.2, CD11c, CD4, CD8a, CD127, CD49b, Ly-6G, Ly-6C, TER-119) ^low, sca-1 ^high, c-kit ^high and was used throughout the manuscript as it best describes the cell population investigated. Granulocyte–macrophage precursors (GMP) were defined as Lin (CD11b, CD19, CD90.2, CD11c, CD4, CD8a, CD127, CD49b, Ly-6G, Ly-6C, TER-119) ^low, c-kit ^high, sca-1 ^low, (CD34/CD16/32) ^high, CD115 ^int/low. Monocyte-dendritic cell precursor (MDP) were defined as Lin (CD11b, CD19, CD90.2, CD11c, CD4, CD8a, CD127, CD49b, Ly-6G, Ly-6C, TER-119) ^low, c-kit ^int/high, sca-1 ^low, (CD34/CD16/32) ^high, CD115 ^high [25 (link)].

Free full text: Click here

Seung H., Wrobel J., Wadle C., Bühler T., Chiang D., Rettkowski J., Cabezas-Wallscheid N., Hechler B., Stachon P., Maier A., Weber C., Wolf D., Duerschmied D., Idzko M., Bode C., von zur Mühlen C., Hilgendorf I, & Heidt T. (2022). P2Y12-dependent activation of hematopoietic stem and progenitor cells promotes emergency hematopoiesis after myocardial infarction. Basic Research in Cardiology, 117(1), 16.

Publication 2022

CD11b (clone M1/70), CD19 (clone 6D5), CD11c (clone N418), CD4 (clone GK1.5), CD8a (clone 53-6.7), clone 2B8

Anti antibodies Antibodies C kit Cd11b Cd90 Cell Clone Dendritic cell First cells Granulocyte Hematopoietic Macrophage Monocyte Mouse Progenitor cells Sca 1 Stem

Corresponding Organization :

Other organizations : Universitäts-Herzzentrum Freiburg-Bad Krozingen, University of Freiburg, Max Planck Institute of Immunobiology and Epigenetics, Inserm, Université de Strasbourg, Établissement Français du Sang, University Medical Centre Mannheim, University Hospital Heidelberg, Medical University of Vienna

Top 5 similar protocols

Variable analysis

independent variables

Incubation of cells with PE-conjugated anti-mouse antibodies directed against CD11b (clone M1/70), CD19 (clone 6D5), CD90.2 (clone 53-2.1), CD11c (clone N418), CD4 (clone GK1.5), CD8a (clone 53-6.7), CD127 (clone A7R34), CD49b (clone DX5), Ly-6G (clone 1A8), Ly-6C (clone HK1.4), TER-119 (clone TER-119)
Staining of cells with antibodies directed against c-kit (BioLegend, clone 2B8), sca-1 (BioLegend, clone D7), CD34 (BD Bioscience, clone RAM34), CD16/32 (BioLegend, clone 93), CD115 (eBioscience, clone AFS98)

dependent variables

Identification and characterization of hematopoietic stem and progenitor cells (LSK cells), granulocyte-macrophage precursors (GMP), and monocyte-dendritic cell precursors (MDP) based on their surface marker expression patterns

control variables

Not explicitly mentioned

controls

No positive or negative controls were specified in the provided information.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!