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Percp cy5.5 conjugated ly 6c

Manufactured by Thermo Fisher Scientific
Sourced in United States

PerCP-Cy5.5 conjugated Ly-6C is a fluorescently labeled antibody that binds to the Ly-6C cell surface antigen. It is commonly used in flow cytometry applications to identify and characterize specific cell populations.

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3 protocols using percp cy5.5 conjugated ly 6c

1

Multicolor Flow Cytometry of Immune Cells

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Single cell suspensions obtained from liver tissues as indicated in 2.4 were stained in FACS buffer with the following anti-mouse antibodies: PE-Cy5 conjugated anti-F4/80 (eBioscience), Alexa Fluor 700 conjugated anti-CD11b (eBioscience), PE-Cy7 conjugated anti-CD45 (eBioscience), FITC conjugated CD4 (BioLegend), BUV395 conjugated CD8a (BD Biosciences), Pacific Blue conjugated CD44 (BioLegend), FITC conjugated F4/80 (BioLegend), FITC conjugated CD3e (BD Biosciences), FITC conjugated Ly-6G (BioLegend), APC-eFluor 780 conjugated CD11c (eBioscience), APC conjugated IFNγ (BioLegend), PerCP-Cy5.5 conjugated Ly-6C (eBioscience) and PE conjugated anti-Ly6G (BioLegend). Acquisition was performed on a LSR Fortessa, LSR Fortessa X-20 or LSRII. Data were analyzed with FlowJo 7.5.5 software (Tree Star).
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2

Quantification of Gastric Macrophages

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Cells were collected from uninured and injured tissue digest, centrifuge at 400×g for 5 min, and were resuspended in PBS with 5% bovine serum albumin. Cells were then incubated with CD16/CD32 FcBlock (BD Biosciences, 553141) for 5 min at 4 °C. Cells were labeled using a 1:100 concentration of antibodies against APC-conjugated F4/80 (Thermo Fisher Scientific, MF48005), FITC-conjugated CD11b (Thermo Fisher Scientific, RM2801), PE-conjugated Ly6G (BD Biosciences, 551461), and PerCP-Cy5.5-conjugated Ly6C (eBiosciences, 45-5932) for 20 min at room temperature in dark and then fixed with fixative medium A (Thermo Fisher Scientific, GAS004) for 15 min at room temperature. Unstained and single-stained cells controls were used as compensation controls. Cells were then washed and resuspended in 500 μL of DPBS containing 0.5% fetal bovine serum and analyzed using FACS Calibur flow cytometer (BD Biosciences). The number of gastric F480+/CD11b+/Ly6Chi/Ly6Gneg macrophages were quantified using FlowJo software according to our previously published protocol14 (link).
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3

Multiparameter Flow Cytometry of Immune Cells

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Cells were stained with the following surface marker antibodies from eBioscience (Waltham, MA, USA): FITC-conjugated CD11b (Clone M1/70), Percp/Cy5.5-conjugated Ly6c (Clone HK1.4), APC-conjugated Gr-1 (Clone RB6-8C5), eFluor 450-conjugated CD45.2 (Cloe 104), PE-conjugated MHC-II (Clone AF6-120.1), PE/Cy7-conjugated F4/80 (Clone, BM8), BV711-conjugated CD45.1 (Clone A20). Cells were acquired on a Fortessa BD Biosciences and analyzed with FlowJo software.
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