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Ly6g pe

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Ly6G-PE is a fluorescently-labeled monoclonal antibody that binds to the Ly6G antigen, which is expressed on the surface of neutrophils. It can be used for the identification and enumeration of neutrophils in flow cytometry applications.

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Lab products found in correlation

Cd11b fitc, by BioLegend (7 mentions) Cd45 percp cy5, by BioLegend (5 mentions) Cd11b apc, by BioLegend (5 mentions) Cd19 fitc, by BioLegend (4 mentions) Cd45 fitc, by BioLegend (4 mentions) Ly6c apc, by BioLegend (4 mentions) Cd45 bv510, by BioLegend (3 mentions) Cd4 pe cy7, by BioLegend (3 mentions) Zombie aqua fixable viability kit, by BioLegend (3 mentions) Cd4 buv805, by BD (3 mentions) Cd3εbuv395, by BD (3 mentions) Cd8a bv785, by BioLegend (3 mentions) Epcam apc af647, by BioLegend (3 mentions) Cd11c buv737, by BD (3 mentions) Nk1.1 buv395, by BD (3 mentions) Ly6c bv785, by BioLegend (3 mentions) Cd11b bv650, by BioLegend (3 mentions) F4 80 bv421 pb, by BioLegend (3 mentions) Cd68 apc cy7, by BioLegend (3 mentions) Anti mouse cd16 cd32 fc block, by BioLegend (3 mentions)

Spelling variants of this product

Anti-Ly6G-PE (22 citations) PE anti-mouse Ly6G (9 citations) Ly6G-PE (1A8, (7 citations) PE-conjugated anti-Ly6G (4 citations) PE/Cy7-conjugated anti-Ly6g (3 citations) PE-conjugated anti-Ly6G antibody (3 citations) PE-Ly6G antibody (2 citations) Anti-Ly6G conjugated to PE (2 citations) PE-conjugated anti-mouse Ly6G antibody (2 citations) α-Ly6G PE-Cy7 (1 citations)

49 protocols using ly6g pe

1

In Vivo Neutrophil Infiltration Assay

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Analysis of in vivo neutrophil infiltration in ear inflammation was performed essentially as described previously(40 (link)). Mice were anaesthetized under isoflurane and approximately 10 μl of 1.5 μM MIP-2 (Peprotech) or PBS was intradermally injected into the pinnae until a blister of 5mm diameter was formed. Pinnae were collected after 4 hours, fixed for 20 min in ice cold 4% paraformaldehyde (PFA), blocked and permeabilised with PBS, 5% BSA, 0.5% Triton X-100 for 3 hours at room temperature and washed three times with PBS. Tissues were stained overnight at 4°C with Ly6G-PE (1A8, Biolegend), CD11b-eFluor660 (M1/70, eBioscience) and VEcadherin-Alexa Fluor 488 (eBioBV13, eBioscience), washed in PBS for 1.5 hours, and compressed between two coverslips for imaging. Neutrophil infiltration was imaged using a 20x multi-immersion objective on a Nikon A1R, with at least 10 fields of view taken per pinna and analysed using ImageJ.
Rynkiewicz N.K., Anderson K.E., Suire S., Collins D.M., Karanasios E., Vadas O., Williams R., Oxley D., Clark J., Stephens L.R, & Hawkins P.T. (2020). Gβγ is a direct regulator of endogenous p101/p110γ and p84/p110γ PI3Kγ complexes in mouse neutrophils. Science signaling, 13(656), eaaz4003.
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2

Corneal Flow Cytometry Analysis

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Mouse eyeballs were collected (n = 5) at 3 days p.i., and the corneas were dissected around the scleral–limbal region. The protocol of corneal flow cytometry has been previously described.34 The gate was set on the CD45+ population. The primary antibodies used in this experiment were CD45‐PercP, CD11b‐fluorescein isothiocyanate and Ly6G‐PE (BioLegend, San Diego, CA, USA).
Gu L., Lin J., Wang Q., Li C., Peng X., Fan Y., Lu C., Lin H., Niu Y., Zhu G, & Zhao G. (2020). Dimethyl itaconate protects against fungal keratitis by activating the Nrf2/HO‐1 signaling pathway. Immunology and Cell Biology, 98(3), 229-241.
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3

Flow Cytometric Analysis of Immune Cells

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For flow cytometric analysis, cells were blocked with PBS containing 1% bovine serum albumin and 0.1% rat IgG (Sigma-Aldrich, St. Louis, USA) for 30 min. After a washing step, cells were stained with SiglecF-PE or -AlexaFluor647 (Clone: E50-2440, BD Pharmingen, San Diego, USA), F4/80-PerCP-Cy5.5 (Clone: BM8), Ly6G-PE (Clone: 1A8), Ly6C-APC-Cy7 (Clone HK1.4) (all BioLegend, San Diego, USA), and CD11b-FITC or -PE-Cy7 (Clone: M1/7; eBioscience, San Diego, USA). For intracellular staining, cells were incubated with fixation and permeabilization buffer (eBioscience) overnight. Cells were stained with rabbit anti-mouse RELMα (Peprotech, Rocky Hill, USA) followed by donkey anti-rabbit IgG AlexaFluor647 (Clone: Poly4064, BioLegend) and CD86-PE (Clone: GL1) and MHCII-APC (Clone: M5/114.15.2, eBioscience) to determine cell activation. The gating strategy used to identify macrophages, monocytes, eosinophils and neutrophils is shown in Figure 1 (Fig. 1).
IFNγ , IL-10, TGFβ and TNF (all eBioscience) as well as CXCL1/KC and CXCL2/MIP-2 (both R&D, Minneapolis, USA) were measured from peritoneal lavage and serum by ELISA according to the manufacturers’ protocols and analyzed using a plate reader (Molecular Devices) with SoftMax Pro 6.
Buerfent B.C., Ajendra J., Stamminger W., Gondorf F., Hoerauf A, & Hübner M.P. (2019). TGFβ depletion does neither modulate acute E. coli-induced inflammatory immune responses nor impair the protective effect by chronic filarial infection. GMS Infectious Diseases, 7, Doc04.
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4

Inflammasome Activation Assay Protocol

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HDM was from Greer Laboratories. Anti-mouse IL-1β (p17) (AF-401-NA) was from R&D. Anti-mouse caspase-1 (p20) (AG-20B-0042) and anti-NLRP3 (AG-20B-0014) antibodies were from Adipogen. Anti-ASC (67824) antibody was from Cell Signaling Technology. Anti-β-actin (66009-1-Ig) was from Proteintech.
Anti-mouse antibodies used for flow cytometry were: CD3-FITC (BD, 553062, 145-2C11), CD19-FITC (eBioscience, 11-0193-82, 1D3), Ly-6G-PE (Biolegend, 127608, 1A8), CD45-PE (eBioscience, 12-0451-81, 30-F11), CD11c-PerCP-Cy5.5 (Biolegend, 117328, N418), CD11b-PerCP-Cy5.5 (BD, 550993, M1/70), CD11b-PE-Cy7 (eBioscience, 25-0112-82, M1/70), CD3e-PE-Cy7 (BD, 552774, 145-2C11), CD19-PE-Cy7 (Biolegend, 115520, 6D5), SiglecF-Alexa Fluor 647 (BD, 562680, E50-2440), MHCII-APC-eFluor 780 (eBioscience, 47-5321-82, M5/114.15.2), Ly-6G-BV421 (Biolegend, 127628, 1A8), CD45-BV510 (Biolegend, 103137, 30-F11), CD11c-BV510 (Biolegend, 117338, N418). Ultrapure LPS was obtained from Invitrogen. Nigericin was obtained from Sigma-Aldrich. RRx-001 (S8405) was from Selleck.
Ma M., Li G., Qi M., Jiang W, & Zhou R. (2021). Inhibition of the Inflammasome Activity of NLRP3 Attenuates HDM-Induced Allergic Asthma. Frontiers in Immunology, 12, 718779.
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5

CpG ODN Encapsulation in PLGA Nanoparticles

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Fully phosphorothioated 3′ Type B CpG 1826 ODN was purchased from Invivogen (sequence: 5′ TCC ATG ACG TTC CTG ACG TT 3′). Poly(vinyl alcohol) (PVA) and chloroform were purchased from Sigma-Aldrich (St. Louis, MO). Research-grade PLGA (50:50, iv .55–.75 dL/g) was purchased from Durect (Pelham, AL). The following fluorochrome-conjugated antibodies were employed: CD11c-PE-Cy7 (BD Biosciences; clone HL3), Ly6C-PerCP-Cyanine5.5 (eBioscience; clone HK1.4), Ly6G-PE (BioLegend; clone 1A8), CD11b-eFluor450 (eBioscience; clone M1/70), CD19-FITC (eBioscience; clone 1D3).
Siefert A.L., Ehrlich A., Corral M.J., Goldsmith-Pestana K., McMahon-Pratt D, & Fahmy T.M. (2016). Immunomodulatory nanoparticles ameliorate disease in the Leishmania (Viannia) panamensis mouse model. Biomaterials, 108, 168-176.
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6

Multiparametric Immune Cell Analysis

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Cells were stained using the following fluorophore conjugated anti-mouse antibodies: WGA-488 (Invitrogen), Ly6G-FitC (clone 1A8, BioLegend), Ly6G-BV510 (clone 1A8, BioLegend), Ly6G-PE (clone 1A8, BioLegend), CD11c-BV605 (clone N418, BioLegend), F4/80-FitC (clone BM8, BioLegend), F4/80-PE (clone BM8, BioLegend), CD11b-PE (clone M1/70, BioLegend), Ly6C-PE Cy7 (clone HK1.4, BioLegend), CD63-APC (clone NVG-2, BioLegend), CD9-APC (clone MZ3, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), and IL-1β-APC (clone NJTEN3, ThermoFisher Scientific). Antibodies were diluted in wash buffer (PBS with 1% BSA and 2 mM EDTA). Cells were stained for 20 minutes at 4°C, washed with wash buffer, fixed for 15 minutes in BD Biosciences cytofix/cytoperm, and permeabilization prior to intracellular staining. ACEA Novocyte was used for flow cytometry, and Novoexpress software was used for subsequent analysis. AMNIS ImageStream was used for imaging flow cytometry and the AMNIS IDEAS software was used to calculate the colocalization coefficient.
Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.
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7

Spinal Cord Injury Immune Cell Analysis

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Spinal cords isolated 1 week after injury and implantation were collected with the bridge and contralateral, but not rostral or caudal tissue, to limit myelin debris. Tissue was digested with 1 U mL−1 liberase at 37°C for 6 minutes in thermomixer (Thermo Scientific) at 1400 RPM. Live cells were detected with a blue fix exclusion dye for 15 minutes at 4 °C. Cells were then incubated for an additional 30 minutes with Ly6G (PE, 1:1000, Biolegend), arginase (FITC, 1:1000, Abcam), CD4 (PECy7, 1:1000, Biolegend), F4/80 (Alexafluor 700, 1:1000, Biolegend), CD11c (PacBlue, 1:1000, Biolegend), GFAP (APC, 1:1000, BD), and CD45 (brilliant violet 510, 1:1000, Biolegend). Cells were then rinsed with saline, fixed with 4% paraformaldehyde for 10 minutes, and rinsed twice more. Samples were analyzed on a MoFlo Astrios flow cytometer using appropriate excitation lasers and emission filters (Beckman Coulter, Brea, CA, USA). Data was analyzed with FlowJo software (FlowJo, Ashland, OR, USA) by investigators blind to the condition.
Dumont C.M., Carlson M.A., Munsell M.K., Ciciriello A.J., Strnadova K., Park J., Cummings B.J., Anderson A.J, & Shea L.D. (2019). Aligned hydrogel tubes guide regeneration following spinal cord injury. Acta biomaterialia, 86, 312-322.
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8

Simultaneous Immune Cell Profiling

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Fresh tumor samples were minced and enzymatically digested with the tumor dissociation kit (Miltenyi #130-096-730) for 40 min at 37°C with agitation. The cell suspension was strained through a 100 μm strainer, spun down and resuspended in 2% FCS/PBS. Cells were blocked for 10 min on ice with anti-mouse CD16/CD32 FC block (Biolegend, 1:100) and stained with Zombie Aqua Fixable Viability Kit (Biolegend, 1:500) and the following antibody cocktails: CD4 BUV805 (BD, 1:100), CD3εBUV395 (BD, 1:20), CD8a BV785 (Biolegend, 1:100), CD45 PerCP Cy5.5 (Biolegend, 1:100), CD19 FITC (Biolegend, 1:100), EpCAM APC/AF647 (Biolegend, 1:200) for acquisition of adaptive immune cells; CD11c BUV737 (BD, 1:30), NK1.1 BUV395 (BD, 1:25), Ly6C BV785 (Biolegend, 1:200), CD11b BV650 (Biolegend, 1:100), F4/80 BV421/PB (Biolegend, 1:30), CD45 PerCP Cy5.5 (Biolegend, 1:100), Ly6G PE (Biolegend, 1:200), CD68 APC-CY7 (Biolegend, 1:20), EpCAM APC/AF647 (Biolegend, 1:200) for acquisition of innate immune cells. Per panel 1,000,000 events were acquired on the BD LSRFortessa. Flow cytometry data was analyzed using FlowJo software (v10.6.2).
Falcomatà C., Bärthel S., Widholz S.A., Schneeweis C., Montero J.J., Toska A., Mir J., Kaltenbacher T., Heetmeyer J., Swietlik J.J., Cheng J.Y., Teodorescu B., Reichert O., Schmitt C., Grabichler K., Coluccio A., Boniolo F., Veltkamp C., Zukowska M., Vargas A.A., Paik W.H., Jesinghaus M., Steiger K., Maresch R., Öllinger R., Ammon T., Baranov O., Robles M.S., Rechenberger J., Kuster B., Meissner F., Reichert M., Flossdorf M., Rad R., Schmidt-Supprian M., Schneider G, & Saur D. (2022). Selective multi-kinase inhibition sensitizes mesenchymal pancreatic cancer to immune checkpoint blockade by remodeling the tumor microenvironment. Nature cancer, 3(3), 318-336.
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9

Multicolor Flow Cytometry of Myeloid Cells

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CD45 APC‐Cy7 (1:100, BioLegend, clone 30‐F11), MHC‐II PE (1:1,000, BioLegend, clone M5), CD11b PE‐Cy7 (1:100, BioLegend, clone M1/70), F4/80 APC (1:50, eBioscience, clone BM8), Ly6G PE (1:50, BioLegend), and 7AAD PerCP (2.5 μl/sample, Sigma).
Beck M.A., Fischer H., Grabner L.M., Groffics T., Winter M., Tangermann S., Meischel T., Zaussinger‐Haas B., Wagner P., Fischer C., Folie C., Arand J., Schöfer C., Ramsahoye B., Lagger S., Machat G., Eisenwort G., Schneider S., Podhornik A., Kothmayer M., Reichart U., Glösmann M., Tamir I., Mildner M., Sheibani‐Tezerji R., Kenner L., Petzelbauer P., Egger G., Sibilia M., Ablasser A, & Seiser C. (2021). DNA hypomethylation leads to cGAS‐induced autoinflammation in the epidermis. The EMBO Journal, 40(22), e108234.
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10

Spleen and Blood Immune Cell Profiling

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To obtain a single cell suspension, spleen was gently disaggregated with a pestle and then with a syringe in 1 ml PBS1x, 2 mM EDTA, after letting debris decant for few minutes, the superior phase was transferred in a new tube. The following procedure was applied to both spleen single cell suspensions and blood samples: after erythrocyte depletion with ACK lysis buffer, 106 cells were suspended in 100 μl RPMI, 10 mM Hepes, 1 mM EDTA, 2% FBS and labeled for 30′ at RT with 1 μg of the following antibodies: CD4-APC (eBioscience, SanDiego, CA, USA), CD8-PeCy7 (eBioscience), CD11b-FITC (eBioscience), B220-PacificBlue (BioLegend, San Diego, CA, USA), Ly-6G-PE (BioLegend), Ly-6C-APCeFluor780 (eBioscience). After labeling cells were washed twice with PBS1x, suspended in 200 μl PBS1x 2 mM EDTA and analyzed with BD LSR II Flow Cytometer (BD Biosciences, San Jose, CA, USA).
Grasso F., Di Meo S., De Luca G., Pasquini L., Rossi S., Boirivant M., Biffoni M., Bignami M, & Di Carlo E. (2015). The MUTYH base excision repair gene protects against inflammation-associated colorectal carcinogenesis. Oncotarget, 6(23), 19671-19684.
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