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Ly6c pe cf594

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Ly6C-PE-CF594 is a fluorescently-conjugated antibody that binds to the Ly6C cell surface antigen. It is commonly used in flow cytometry applications for the identification and analysis of Ly6C-positive cell populations.

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Lab products found in correlation

Cd11b apc, by Thermo Fisher Scientific (2 mentions) Cd4 apc, by Thermo Fisher Scientific (2 mentions) Cd8 fitc, by Thermo Fisher Scientific (2 mentions) Cd3 percp cy5, by Thermo Fisher Scientific (2 mentions) Pd1 pecf594, by BD (2 mentions) Ly6g percp cy5, by BD (2 mentions) Fcr blocker, by Thermo Fisher Scientific (2 mentions) Anti gr1 rb6, by BD (2 mentions) Mouse tumor cell dissociation kit, by Miltenyi Biotec (2 mentions) Cytoflex s flow cytometer, by Beckman Coulter (1 mentions) Cd45 vf450, by Cytek Biosciences (1 mentions) Cd11b apc cy7, by Cytek Biosciences (1 mentions) Lsrfortessa flow cytometer, by BD (1 mentions) Cd45 apc cy7, by BD (1 mentions) Cd4 percp, by BD (1 mentions) Neural tissue dissociation kit, by Miltenyi Biotec (1 mentions) Red blood cell lysis solution, by Miltenyi Biotec (1 mentions) Cd19 pe cf594, by BD (1 mentions) Cell staining buffer, by BD (1 mentions) Cd11b bv421, by BD (1 mentions)

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Ly6C-PE (17 citations)

4 protocols using ly6c pe cf594

1

Quantitative Analysis of Myeloid-Derived Suppressor Cells and T-Cells in Tumor Samples

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To characterize and quantify MDSCs by FACS, cells were incubated with FcR blocker (eBioscience), then stained with Ly6C-PE-CF594 (BD Biosciences), Ly6G-PerCPCy5.5 (BD Bioscience), CD11b-APC (eBioscience) and/or anti-GR1 (RB6, BD Biosciences). MDSCs were identified as GR1+CD11b+ cells, granulocytic MDSCs as Ly6GhighLy6Clow or −CD11b+, monocytic MDSCs as Ly6ChighLy6GlowCD11b+. To determine absolute cell numbers counting beads (BD Biosciences) were added before FACS acquisition. T-cells were analyzed with an antibody combination of CD3-PerCPCy5.5, CD4-APC, CD8-FITC (all eBioscience) and PD1-PE-CF594 (BD Biosciences). Please also see Supplementary Table 2 for antibody use information.
Tumor infiltrating MDSCs and T-cells were quantitated by enzymatic digestion of the dissected tumors (Mouse tumor cell dissociation kit, Miltenyi Biotec), followed by FACS staining with the above antibody combinations. Staining for the common leukocyte marker CD45 was also included to facilitate the distinction of leukocytes from tumor cells and other stromal cells.
Welte T., Kim I.S., Tian L., Gao X., Wang H., Li J., Holdman X.B., Herschkowitz J.I., Pond A., Xie G., Kurley S., Nguyen T., Liao L., Dobrolecki L.E., Mo Q., Edwards D.P., Huang S., Xin L., Xu J., Li Y., Lewis M.T., Wang T., Westbrook T.F., Rosen J.M, & Zhang X.H. (2016). Oncogenic mTOR signaling recruits myeloid-derived suppressor cells to promote tumor initiation. Nature cell biology, 18(6), 632-644.
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2

Multiparametric Immune Cell Phenotyping

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Following this blocking step, cells were re-suspended in the following cocktail of fluorescently conjugated antibodies: CD45-vf450 (Tonbo Biosciences), CD11b APC-Cy7 (Tonbo Biosciences), Ly6G-PeCy (Tonbo Biosciences) and Ly6C-PeCF594 (BD Biosciences) and incubated for 30 minutes at room temperature. After staining, cells were washed twice and re-suspended in 300ul FACS buffer for analysis on a Cytoflex S Flow Cytometer (Beckmann Coulter). Data files were analyzed using FlowJo (TreeStar). The representative gating strategy used is given in Supplementary Figure 2. Positive and negative populations were defined using fluorescence minus one (FMO) controls.
Roy-O’Reilly M.A., Ahnstedt H., Spychala M.S., Munshi Y., Aronowski J., Sansing L.H, & McCullough L.D. (2020). Aging exacerbates neutrophil pathogenicity in ischemic stroke. Aging (Albany NY), 12(1), 436-461.
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3

Quantitative Analysis of Myeloid-Derived Suppressor Cells and T-Cells in Tumor Samples

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To characterize and quantify MDSCs by FACS, cells were incubated with FcR blocker (eBioscience), then stained with Ly6C-PE-CF594 (BD Biosciences), Ly6G-PerCPCy5.5 (BD Bioscience), CD11b-APC (eBioscience) and/or anti-GR1 (RB6, BD Biosciences). MDSCs were identified as GR1+CD11b+ cells, granulocytic MDSCs as Ly6GhighLy6Clow or −CD11b+, monocytic MDSCs as Ly6ChighLy6GlowCD11b+. To determine absolute cell numbers counting beads (BD Biosciences) were added before FACS acquisition. T-cells were analyzed with an antibody combination of CD3-PerCPCy5.5, CD4-APC, CD8-FITC (all eBioscience) and PD1-PE-CF594 (BD Biosciences). Please also see Supplementary Table 2 for antibody use information.
Tumor infiltrating MDSCs and T-cells were quantitated by enzymatic digestion of the dissected tumors (Mouse tumor cell dissociation kit, Miltenyi Biotec), followed by FACS staining with the above antibody combinations. Staining for the common leukocyte marker CD45 was also included to facilitate the distinction of leukocytes from tumor cells and other stromal cells.
Welte T., Kim I.S., Tian L., Gao X., Wang H., Li J., Holdman X.B., Herschkowitz J.I., Pond A., Xie G., Kurley S., Nguyen T., Liao L., Dobrolecki L.E., Mo Q., Edwards D.P., Huang S., Xin L., Xu J., Li Y., Lewis M.T., Wang T., Westbrook T.F., Rosen J.M, & Zhang X.H. (2016). Oncogenic mTOR signaling recruits myeloid-derived suppressor cells to promote tumor initiation. Nature cell biology, 18(6), 632-644.
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4

Mouse Brain Cell Isolation and Flow Cytometry

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PBS-perfused mouse cerebra at P7 were digested with a Neural Tissue Dissociation Kit (Miltenyi Biotec), and cell debris and residual red blood cells were removed using the Debris Removal Solution (Miltenyi Biotec) and Red Blood Cell Lysis Solution (Miltenyi Biotec) according to the manufacturer’s instructions, respectively. The cells were stained with antibodies against mouse antigens of CD45 APC-Cy7 (BD Biosciences, 557659), CD4 PerCP (BD Biosciences, 553052), CD8a BV650 (BD Biosciences, 563234), CD3 MolCpx BV421 (BD Biosciences, 564008), CD19 PE-CF594 (BD Biosciences, 562291), CD49b APC (BD Biosciences, 560628), Ly-6G BUV395 (BD Biosciences, 565964), Ly-6C PE-CF594 (BD Biosciences, 562728), CD16/CD32 PE-Cy7 (BD Biosciences, 560829), F4/80 Alexa Fluor 647 (BD Biosciences, 565853), or CD11b BV421 (BD Biosciences, 562605) at room temperature for 20 minutes, then washed twice with Cell Staining Buffer (BD Biosciences, 554657). The stained cells were resuspended with 300 μL of Cell Staining Buffer and applied to a BD LSRFortessa Flow Cytometer (BD Biosciences).
Zhou Y.P., Mei M.J., Wang X.Z., Huang S.N., Chen L., Zhang M., Li X.Y., Qin H.B., Dong X., Cheng S., Wen L., Yang B., An X.F., He A.D., Zhang B., Zeng W.B., Li X.J., Lu Y., Li H.C., Li H., Zou W.G., Redwood A.J., Rayner S., Cheng H., McVoy M.A., Tang Q., Britt W.J., Zhou X., Jiang X, & Luo M.H. (2022). A congenital CMV infection model for follow-up studies of neurodevelopmental disorders, neuroimaging abnormalities, and treatment. JCI Insight, 7(1), e152551.
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