Ly6g percp cy5

Manufactured by Thermo Fisher Scientific

Ly6G-PerCP-Cy5.5 is a fluorescently-labeled antibody that binds to the Ly6G antigen. It is commonly used in flow cytometry applications to identify and quantify granulocytes, particularly neutrophils, in biological samples.

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Lab products found in correlation

Lsrii flow cytometer, by BD (3 mentions) Cd11c pe txr, by R&D Systems (2 mentions) Cd64 bv421, by BioLegend (1 mentions) Anti cd40 pe, by BD (1 mentions) Anti mhcii fitc, by BD (1 mentions) Anti cd45 v450, by BD (1 mentions) Anti cd86 pe, by BD (1 mentions) Anti f4 80 apc, by BD (1 mentions) Anti ly6c pe cy7, by BD (1 mentions) Anti cd11b apc cy7, by BD (1 mentions) Ccr2 apc, by Thermo Fisher Scientific (1 mentions) 70 m cell strainer, by BD (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Facsvantage se, by BD (1 mentions) Cd45 apc, by Thermo Fisher Scientific (1 mentions) Fc receptor blocking antibody, by Thermo Fisher Scientific (1 mentions) Cd11c apc efluor780, by Thermo Fisher Scientific (1 mentions) Cd11b fitc, by Thermo Fisher Scientific (1 mentions) F4 80 pe, by Thermo Fisher Scientific (1 mentions) F4 80 efluor450, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PerCP-Cy5.5 (62 citations) Anti-ly6G-PerCP-Cy5.5 (2 citations)

5 protocols using ly6g percp cy5

Monocyte and DC Recruitment in Mice

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To analyze monocyte and DC recruitment in response to CpG, 20 µl of a 100 µg/ml ODN1864 solution was injected into the footpad of C57BL/6 J mice. At the indicated time intervals post injection, blood samples were collected through the tail vein followed by ACK (Thermo Fisher Scientific) mediated red blood cell lysis. Alternatively, mice were euthanized and the DLNs were dissected. Single cell suspensions of DLN were prepared through incubation with collagenase type IV (Sigma-Aldrich) and passed through a 70 µm cell strainer (BD Falcon).
After treatment with 2.4G2 Ab for 5 min to block the FcR, PBMCs or lymph node cell suspensions were stained with the following anti-mouse Abs: anti-CD45-V450, anti- MHCII-FITC, anti-CD86-PE, anti-CD40- PE, anti-CD64-APC, anti-F4/80-APC, anti-Ly6C-PE-Cy7, anti-CD11b-APC-Cy7 (all BD Biosciences), Ly6G-PerCP-Cy5.5, CCR2-APC (eBioscience), CD64-BV421 (BioLegend) and CD11c-PE-TxR (R & D systems). Death cells were identified by staining with aqua life/dead stain (Invitrogen). Fluorescent events were acquired using an LSRII flow cytometer (BD Biosciences) and analyzed using FlowJo software. After exclusion of dead cells, granulocytes were identified as CD45+ CD11b+ Ly6C+ Ly6G+ cells and Monocytes as CD45+ Ly6C^hi CD11b^hi Ly6G- cells. Following exclusion of granulocytes and Monocytes in the LN, DCs were subdivided into MHCII^hi DCs and MHCII^int DCs.

De Koker S., Van Hoecke L., De Beuckelaer A., Roose K., Deswarte K., Willart M.A., Bogaert P., Naessens T., De Geest B.G., Saelens X., Lambrecht B.N, & Grooten J. (2017). Inflammatory monocytes regulate Th1 oriented immunity to CpG adjuvanted protein vaccines through production of IL-12. Scientific Reports, 7, 5986.

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Characterizing Lung Immune Cell Responses Post-Irradiation

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BAL was performed prior to lung resection at 1, 8, 12, and 18 weeks after irradiation. Cells were loaded onto slides using a cytospin centrifuge and stained using a DiffQuik staining kit (IMEB Inc., San Marcos, CA,). Differential cell counts of leukocyte subsets were performed by counting at least 300 nucleated cells.²⁶Following collection of BAL fluids, the same mice provided the lungs for flow cytometry studies. Lungs were digested with 0.4 mg/mL collagenase IV and red blood cells were lysed. Lung single-cell suspensions were incubated with Fc receptor-blocking antibody (eBioscience, San Diego, CA) prior to staining. For morphological characterization of leukocytes, CD45⁺ cells were sorted by fluorescence-activated cell sorting (FACS) using a BD FACSVantage SE. CD45⁺ cell subsets were gated according to cell size and granularity. Cell subsets obtained from each gate were spun onto slides using a cytospin, and stained using a DiffQuik staining kit. To determine immunophenotype, cells were immunostained using a 5-color fluorophore combination of antibodies consisting of CD45-APC, CD11b-FITC, F4/80-PE, CD11c-APC-eFluor780, and Ly6G-PerCp-Cy5.5 (eBioscience). Fixable viability dye eFluor450 was used to exclude dead cells. Cells were analyzed by flow cytometry using a BD LSR II flow cytometer followed by analysis on FlowJo v10 software.

Abernathy L.M., Fountain M.D., Rothstein S.E., David J.M., Yunker C.K., Rakowski J., Lonardo F., Joiner M.C, & Hillman G.G. (2015). Soy Isoflavones Promote Radioprotection of Normal Lung Tissue by Inhibition of Radiation-Induced Activation of Macrophages and Neutrophils. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 10(12), 1703-1712.

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Flow Cytometry of Mouse Immune Cells

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For flow cytometry, diaphragms were digested for 1 hour in 5 mL 0.2% collagenase II (MP Biomedicals) at 37°C. Collagenase was inactivated with addition of FACS buffer (2% FCS in dPBS with 2 mM EDTA). Tissue was filtered through a 70 μm, strainer and washed (centrifugation at 400g at 4°C for 5 minutes). Lysis buffer was added for 1 minute to remove red blood cells and washed 2x with FACS buffer. Cell viability was determined using Fixable Viability Dye eFluor 780 (eBioscience, UK) at a dilution of 1/1000 in FACS buffer for 20 minutes at 4°C. Cells were washed and incubated with fluorophore‐conjugated antibodies at 1/200 dilution in Fc block (anti‐mouse CD16/32, BioLegend, in FACS buffer) for 20 minutes at 4°C. Antibodies used were CD45‐PE (eBioscience, clone 30‐F11), CD11b‐APC (BD pharminogen, clone M1/70), Ly6G‐PerCPCy5.5 (eBioscience, clone1A8‐Ly6g), F4/80‐eFluor 450 (eBioscience, clone BM8), NOS‐PECy7 (eBioscience, clone CXNFT), and CD206‐AF 700 (eBioscience, clone C068C2). Data were acquired using an LSRII flow cytometer (BD Biosciences) and analysed using FlowJo software (TreeStar version 7.6.5).

Cunningham M.E., Meehan G.R., Robinson S., Yao D., McGonigal R, & Willison H.J. (2020). Perisynaptic Schwann cells phagocytose nerve terminal debris in a mouse model of Guillain‐Barré syndrome. Journal of the Peripheral Nervous System, 25(2), 143-151.

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Comprehensive Lung Cell Profiling by Flow Cytometry

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At 4 and 7 dpi, we isolated lung cells as previously described [38 ] and assessed lung cellular composition. Cells were stained at 4°C for 20 min with fluorescently labeled antibodies against the following cell surface markers purchased from eBiosciences unless otherwise noted: Siglec F PE (clone IRNM44N), CD11b APC (clone M1/70), CD11c Efluor 450 (clone N418), Ly6C PE-Cy7 (clone HK1.4), Ly6G PerCP-Cy5.5 (Tonbo, clone 1A8), CD45.2 FITC (clone 104) [Stain 1]; TCRαβ FITC (clone H57-597), TCRγδ APC (clone eBioGL3), CD4 PE-Cy7 (clone GK1.5), CD8 Efluor 450 (clone 53–6.7), DX5 PE (clone DX5), CD45.2 PerCP-Cy5.5 (clone 104) [Stain 2]. The cells were fixed with 1% formalin and analyzed on the LSR II flow cytometer (BD Biosciences). CD45.2-gated hematopoietic-derived leukocyte populations were characterized as follows: inflammatory monocytes (IM; Siglec F⁻ CD11b^high CD11c^low Ly6C⁺), neutrophils (Siglec F⁻ CD11b^high CD11c^low Ly6G⁺), alveolar macrophages (AM; Siglec F^high CD11b^int), eosinophils (Siglec F^high CD11b^high), NK cells (αβ TCR⁻ DX5⁺), CD4 T cells (αβ TCR⁺ CD4⁺), CD8 T cells (αβ TCR⁺ CD8⁺), and γδ T cells (αβ TCR⁻ γδ TCR⁺).

Latha K., Patel Y., Rao S, & Watford W.T. (2022). The Influenza-Induced Pulmonary Inflammatory Exudate in Susceptible Tpl2-Deficient Mice Is Dictated by Type I IFN Signaling. Inflammation, 46(1), 322-341.

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Characterizing Antigen Uptake by Monocytes and DCs

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To evaluate antigen uptake by Ly6C^hi monocytes, MHCII^hi and MHCII^int DCs in the lymph node, Alexa647-labbeled OVA (100 µg/ml) was injected into the footpad of mice mixed with ODN1864 (100 µg/ml). Popliteal lymph nodes were dissected at the indicated time intervals and single cell suspensions were stained with aqua life/dead, Fc block, CD45-MHCII-FITC, Ly6-PE-Cy7, CD11b-APC-Cy7 (BD-biosciences), CD11c-PE-TxR (R & D systems) and Ly6G-PerCP-Cy5.5 (eBioscience). To determine the OVA-Alexa647 positive monocyte/DC gate, mice were injected with a mixture of unlabeled OVA (100 µg/ml) and ODN1864 (100 µg/ml).

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