Flow Cytometry
Selection of Specific Markers on Mouse Cell Surface
Source: Elabscience®Published: Jan 17,2024
F4/80, NK1.1, CD90.1/CD90.2, CD45.1/CD45.2,these specific Markers of mouse cell surface are often mentioned in the literature.Do you feel confused to these markers? Same cell model, why do some mice choose one marker and some mice choose another ? How to choose these markers? Why we need to use CD16/32 as FCR block and how to use it? The answers of these questions are following !
F4/80-Markers of mature mouse macrophages
F4/80 is a 160 kD glycoprotein. It is characterized as a member of the epidermal growth factor (EGF)-transmembrane 7 (TM7) family. F4/80, also known as EMR1 or Ly71, has been widely used as a murine macrophage marker, which is expressed on the majority of tissue macrophages including peritoneal macrophages, macrophages in lung, gut, thymus and red pulp of spleen (but not on the macrophages located in T cell areas of the spleen, lymph node and Peyer's patch), Kuffer cells, Langerhans cells, and bone marrow stromal cells. F4/80 has also been shown on a subset of dendritic cells. The biological ligand of F4/80 has not been identified, but it has been reported that F4/80 is required for induction of CD8+ T cells-mediated peripheral tolerance.
CD16/32 - Cell surface Fc receptor
CD16 is low affinity IgG Fc receptor III (FcR III) and CD32 is FcR II. CD16/CD32 are expressed on B cells, monocytes/macrophages, NK cells, granulocytes, mast cells, and dendritic cells. The Fc receptors bind antibody-antigen immune complexes and mediate adaptive immune responses.
CD49b and NK1.1 in mouse NK cells
DX5 antigen has been recently characterized as CD49b. It is a 150 kD integrin α chain also known as α2 integrin, VLA-2 α chain, and integrin α2 chain. CD49b non-covalently associates with CD29 (β1 integrin) to form the CD49b/CD29 complex known as VLA-2, a receptor for collagen and laminin. CD49b is expressed on platelets, the majority of NK cells, NKT cells, and a small subset of CD8+ T cells (this population can be significantly increased following viral infection). DX5 is used for the identification and isolation of NK cells, and is especially useful for identifying NK cells in mice lacking the NK1.1 antigen.
NK-1.1 surface antigen, also known as CD161b/CD161c and Ly-55, is encoded by the NKR-P1B/NKR-P1C gene. It is expressed on NK cells and NK-T cells in some mouse strains, including C57BL/6, FVB/N, and NZB, but not AKR, BALB/c, CBA/J, C3H, DBA/1, DBA/2, NOD, SJL, and 129. Expression of NKR-P1C antigen has been correlated with lysis of tumor cells in vitro and rejection of bone marrow allografts in vivo. NK-1.1 has also been shown to play a role in NK cell activation, IFN-γ production, and cytotoxic granule release. NK-1.1 and DX5 are commonly used as mouse NK cell markers.
Differences between Mouse CD90 ,CD90.1 and CD90.2
CD90 is a 25-35 kD immunoglobulin superfamily member, also known as Thy-1. It is expressed on hematopoietic stem cells and neurons, all thymocytes, and peripheral T cells, but not on B cells. CD90 is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein involved in signal transduction. CD90 is involved in costimulation of lymphocyte proliferation and activation, and hematopoietic stem cell differentiation. CD90 has been shown to interact with CD45.
CD90.1 is a 25-35 kD immunoglobulin superfamily member, also known as Thy-1. It is expressed on hematopoietic stem cells and neurons, all thymocytes, and peripheral T cells. CD90.1 is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein involved in signal transduction. CD90.1 is involved in costimulation of lymphocyte proliferation and activation, and hematopoietic stem cell differentiation. CD90.1 has been shown to interact with CD45.
CD90.2 is a 25-35 kD immunoglobulin superfamily member also known as Thy1.2. It is expressed on hematopoietic stem cells and neurons, all thymocytes, and peripheral T cells in Thy1.2 bearing mouse strains (Balb/c, CBA/J, C3H/He, C57BL/-, DBA, NZB/-). CD90.2 is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein involved in signal transduction. CD90.2 is involved in costimulation of lymphocyte proliferation and induction of hematopoietic stem cells differentiation. CD90.2 has been shown to interact with CD45. The 30H12 antibody has been reported to induce Ca2+ flux in thymocytes and, in combination with antibody against the CD3/TCR complex, promote thymocyte apoptosis and inhibit CD3-mediated proliferative responses of mature T lymphocytes.
The difference between Mouse CD45, CD45.1 and CD45.2.
CD45 is a 180-240 kD glycoprotein also known as the leukocyte common antigen (LCA), T200, or Ly-5. It is a member of the protein tyrosine phosphatase (PTP) family, expressed on all hematopoietic cells except mature erythrocytes and platelets. There are different isoforms of CD45 that arise from alternative splicing of exons 4, 5, and 6, which encode A, B, and C determinants, respectively. CD45 plays a key role in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation state of the cell as well as cell type. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.
CD45.1 is an alloantigen of CD45, expressed by Ly5.1 bearing mouse strains (e.g., RIII, SJL/J, STS/A, DA). CD45, a member of the protein tyrosine phosphatase (PTP) family, is a 180-240 kD glycoprotein expressed on all hematopoietic cells except mature erythrocytes and platelets. There are multiple isoforms in mice that play key roles in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation states of the cell as well as specific cell types. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.
CD45.2 is an alloantigen of CD45, expressed by Ly5.2 bearing mouse strains (e.g., A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C57BR, C57L, C58, DBA/1, DBA/2, NZB, SWR, 129). CD45, a member of the protein tyrosine phosphatase (PTP) family, is a 180-240 kD glycoprotein expressed on all hematopoietic cells except mature erythrocytes and platelets. There are multiple isoforms in the mouse that play key roles in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation states of the cell as well as specific cell type. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1.