Ly6g apc cy7

Manufactured by BioLegend

Sourced in United States

Ly6G-APC-Cy7 is a fluorophore-conjugated antibody that binds to the Ly6G antigen, which is expressed on the surface of neutrophils. This antibody can be used for the identification and analysis of neutrophils in flow cytometry applications.

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Lab products found in correlation

F4 80 pe cy7, by BioLegend (4 mentions) Ly6c pe cy7, by BioLegend (4 mentions) Ly6c pe, by BioLegend (3 mentions) Cd11b percp cy5, by BioLegend (3 mentions) Facscanto 2, by BD (3 mentions) Lsrii flow cytometer, by BD (3 mentions) Cd11b apc, by BioLegend (3 mentions) Cd11c pe cy7, by BioLegend (2 mentions) Cd11b fitc, by BioLegend (2 mentions) Foxp3 pe, by Thermo Fisher Scientific (2 mentions) Anti mouse ly6c apc, by BioLegend (2 mentions) Streptavidin pe cy5, by Thermo Fisher Scientific (2 mentions) Cd11b biotin, by Thermo Fisher Scientific (2 mentions) Il 17 alexa fluor 647, by Thermo Fisher Scientific (2 mentions) Siglecf pe, by BD (2 mentions) Cd11b pe cy7, by BioLegend (2 mentions) Cd4 apc, by BioLegend (2 mentions) Cd4 apc cy7, by BioLegend (2 mentions) Cd45 alexa fluor 700, by BioLegend (2 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

APC-Cy7 (106 citations) Ly6G-APC (33 citations) Anti-Ly6G-APC/Cy7 (10 citations) APC-Cy7–conjugated anti-Ly6G (6 citations) Anti-Ly6G-APC/Cy7 (1A8), (2 citations) APC/Cy7-anti-Ly6G mAb (clone 1A8, (2 citations) Anti-Ly6G-APC/Cy7 (clone 1A8) (2 citations)

28 protocols using ly6g apc cy7

Flow Cytometric Analysis of Muscle Inflammation

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For flow cytometric muscle analysis, mice were euthanized by CO2 asphyxiation 7 days after injury to analyze cellular inflammation in the muscle. Tissue was minced, digested in 1mg/mL collagenase IA for 45 min at 37°C, filtered through membranes with 40μm pore size, and resuspended in 3% FBS for immunostaining. Cell suspensions were immunostained for 30 min on ice followed by fixation in 2% PFA for 10 min and addition of CountBrightTM Absolute Counting Beads. The following antibody panel was used: MerTK-PE (clone 108928; R&D Systems), CCR7-PE/Cy7 (clone 4B12; BioLegend), CD3-FITC (17A2; BioLegend), CD25-PerCP/Cy5.5 (clone PC61; BioLegend), Ly6C-APC (clone HK1.4, BioLegend), Ly6G-APC/Cy7 (clone 1A8; BioLegend), CD11c-BV421 (clone N418; BioLegend), CD11b-BV510 (clone M1/70; BioLegend), CD206-BV605 (clone C068C2; BioLegend), CD64-BV711 (clone X54-5/7.1; BioLegend), and CD4-BV785 (clone GK1.5; BioLegend). Samples were run on a BD FACS Aria IIIu cytometer and data was analyzed using FlowJo software. Cells were immunophenotyped according to the following gating scheme: macrophage, MerTK+CD64+; dendritic cell, NOT(MerTK+CD64+)CD11c+; monocyte, NOT(MerTK+CD64+)CD11c-CD11b+SSClo; neutrophil, Ly6G+SSChi; T lymphocyte, CD3+; helper T lymphocyte, CD3+CD4+; regulatory T lymphocyte (Treg), CD3+CD4+CD25+.

Krieger J.R., Sok M.C., Turner T.C, & Botchwey E.A. (2018). Delivery of Immunomodulatory Microparticles in a Murine Model of Rotator Cuff Tear. MRS advances, 3(24), 1341-1346.

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Comprehensive Tumor Immune Profiling

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Tumor cells isolated from mice were thawed and stained with LIVE/DEAD Fixable Violet Dead Staining Kit (ThermoFisher). Subsequently, the cells were divided and stained with cocktails of fluorochrome-conjugated monoclonal antibodies: CD3 PE-CF594, CD19 PE-CF594, CD49b PE-CF594 (all from BD Biosciences), CD45 BV605, CD11b PerCP-Cy5.5, CD11c BV650, F4/80 AlexaFluor 700, Ly6C PE, Ly6G APC-Cy7, MHC II FITC, CD80 PE-Cy7 (all from Biolegend) for myeloid cell identification and CD45 BV605, CD3 BV650, CD4 FITC, CD8 APC-Fire, CD25 PE, CD44 PE-Cy7, CD62L PerCP-Cy5.5 (all from BioLegend) for lymphocytes identification. Then, the cells were fixed using FoxP3 Fixation Permeabilization Staining Kit (eBioscience). Tumor cells stained with myeloid or lymphocyte cocktail were additionally incubated with anti-CD206 APC (BioLegend) or FoxP3 APC (eBioscience) antibodies, respectively. The analysis was performed using FACSFortessa flow cytometer with Diva software (Becton Dickinson).

Rossowska J., Anger N., Szczygieł A., Mierzejewska J, & Pajtasz-Piasecka E. (2018). Reprogramming the murine colon cancer microenvironment using lentivectors encoding shRNA against IL-10 as a component of a potent DC-based chemoimmunotherapy. Journal of Experimental & Clinical Cancer Research : CR, 37, 126.

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Flow Cytometry Panel for Immune Cell Profiling

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Flow cytometry antibodies: B220/CD45R-e450, CD3-e780, CD11b-APC, CD11c-e780, CD19-e780, CD49b-e780 CD86-Alexa Fluor 488, NK1.1-e780 from eBioscience. CD8α-PECF594, CD40-PE from BD Pharmingen. B220-BV650, CD11b-BV711, CD11c-BV421, CD40-FITC, CD80-BV605, CD86-A700, F4/80-PE-Cy7, Ly6C-BV570, Ly6G-APC-Cy7, MHC II-PerCP-Cy5.5 from Biolegend. PDCA-1-PE, FcR block from Miltenyi Biotec. The S9.6 monoclonal antibody against RNA:DNA hybrids was purified from hybridoma cell line HB-8730 (ATCC-LGC Promochem) supernatant using a Protein A/G column as previously described (Pohjoismaki et al, 2010 (link)).

Rigby R.E., Webb L.M., Mackenzie K.J., Li Y., Leitch A., Reijns M.A., Lundie R.J., Revuelta A., Davidson D.J., Diebold S., Modis Y., MacDonald A.S, & Jackson A.P. (2014). RNA:DNA hybrids are a novel molecular pattern sensed by TLR9. The EMBO Journal, 33(6), 542-558.

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Multiparameter Immunophenotyping of Cells

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For flow cytometric analysis of surface markers and intracellular cytokines, single cell suspensions of lungs, spleens, or cell cultures were stained with optimal concentrations of the following specific antibodies: CD45-V450, CD4-V500, CD8-V450, and CD62L-APC from BD Biosciences, CD3-PerCP-Cy5.5, CD4-BV510, CD4-PE-Cy7, CD8a-FITC, CD44-FITC, CD19-PE, CD80-AF488, CD86-APC, Ly6G-APC-Cy7, CD11c-PE-Cy7, NK1.1-PE-Cy7, TNFα-Pacific Blue, IFNγ-PerCP-Cy5.5, IL-17A-PerCP-Cy5.5, IL-2-PE-Cy7, and IL-10-PE from BioLegend and CD90.2-eFluor780 from eBioscience. Data were acquired on a FacsCantoII^® flow cytometer (BD Biosciences) equipped with a 405, 488, and 633 nm laser and analyzed with the FCS Express software (DeNovo™ Software).

Blank J., Eggers L., Behrends J., Jacobs T, & Schneider B.E. (2016). One Episode of Self-Resolving Plasmodium yoelii Infection Transiently Exacerbates Chronic Mycobacterium tuberculosis Infection. Frontiers in Microbiology, 7, 152.

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Flow Cytometry Analysis of Immune Cells

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Single cell suspensions of blood (by cardiac puncture) were collected from NRL-CV, NRL and wild-type adult male and female mice and stained for flow cytometry. Antibodies used: CD3-BV785 (17A2, BD Biosciences, cat 564010), CD45-BV711 (30-F11, Biolegend, cat 103147), CD4-PE-Cy7 (RM4–4, Biolegend, cat 116016), CD8-BUV395 (53–6.7, BD Bioscience, cat 563786), CD19-BV605 (6D5, Biolegend, cat 115540), NKp46-BUV737 (29A1.4, BD Bioscience, cat 612805), LY6G-APC-Cy7 (1A8, Biolegend, cat 127623), CD11b-BV650 (M1/70, Biolegend, cat 101259), SiglecF-AF647 (E50–2440, BD Bioscience, cat 562680), CD64-PerCPCy5.5 (X54–5/7.1, Biolegend, cat 139308), TCRγδ-FITC (GL3, Biolegend, cat 118105), CD11c-BV510 (N418, Biolegend, cat 117353) and counterstained with DAPI (ThermoFisher, cat D1306). Samples were acquired on a BD Fortessa instrument. Data were analysed using FlowJo v10.8.1. For cell gating, RFP+ cells were subgated out of CD45+DAPI− single and the immune subsets were subgated out of the RFP+ gate (see Table 1 and Supplementary Figure 4a).

Dzien P., Raffo Iraolagoitia X., May S., Stevenson D., McGarry L., Soloviev D., Brown G., Nixon C., Kapeni C., De La Roche M., Blyth K., Lyons S., Bird T., Strathdee D., Fruhwirth G., Carlin L, & Lewis D. (2024). Multi-scale in vivo imaging of tumour development using a germline conditional triple-reporter system. Research Square.

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Boronic Acid-Based Glucose-Responsive Hydrogels

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N-isopropylmethacrylamide (NIPMAAm), 4-(2-acrylamidoethylcarbamoyl)-3-fluorophenylboronic acid (AmECFPBA), N,N′-methylenebisacrylamide (MBAAm), 2,2′-azobisisobutyronitrile (AIBN), FITC-labeled bovine insulin, and all organic solvents (acetone, dichloromethane, ethanol, hexane, toluene, and DMSO) were all purchased from Wako Pure Chemical Industries. NIPMAAm was recrystallized in hexane and then dried in vacuo overnight before use. Antibodies CD45-PE/Cy7, CD11b-FITC, B220-APC, CD3ε-PE, and Ly6G-APC/Cy7 for flow cytometry were purchased from BioLegend. All other reagents were purchased from Sigma-Aldrich or Nacalai Tesque and used as received, unless otherwise noted.

Matsumoto A., Tanaka M., Matsumoto H., Ochi K., Moro-oka Y., Kuwata H., Yamada H., Shirakawa I., Miyazawa T., Ishii H., Kataoka K., Ogawa Y., Miyahara Y, & Suganami T. (2017). Synthetic “smart gel” provides glucose-responsive insulin delivery in diabetic mice. Science Advances, 3(11), eaaq0723.

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Characterizing Autoimmune Neuroinflammation

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Myelin oligodendrocyte glycoprotein 35–55 (MOG_35–55) peptide (MEVGWYRSPFSRVVHLYRNGK) and myelin basic protein (MBP) Ac_1–11 peptide (Ac-ASQKRPSQRSK) were generated by the Protein Core Laboratory of the Blood Research Institute, BloodCenter of Wisconsin. KYC was synthesized by Biomatik (Wilmington, DE). The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780 CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse IFN-γ-PE was purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). The SMI-32 antibody, which detects nonphosphorylated neurofilament-H was purchased from Covance (Emeryville, CA). Streptavidin Alexa 405 and goat anti-mouse Alex 456 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-MPO heavy chain was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti-rabbit IgG (H + L)-HRP was purchased from Jackson ImmunoResearch (West Grove, PA). Anti-β-actin-HRP was purchased from Sigma-Aldrich (St. Louis, MO). For all experiments, the mice were age (6–8 wk) and gender matched with both sexes being utilized.

Zhang H., Ray A., Miller N.M., Hartwig D., Pritchard KA J.r, & Dittel B.N. (2015). Inhibition of Myeloperoxidase at the Peak of Experimental Autoimmune Encephalomyelitis Restores Blood-Brain-Barrier Integrity and Ameliorates Disease Severity. Journal of neurochemistry, 136(4), 826-836.

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Flow Cytometry Analysis of Immune Cells

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Ketamine (10 mg/kg/mouse) or saline solution for control mice was administered via intra-peritoneal injection [33] (link). After 72 h, animals were euthanized and peripheral blood, bone marrow, spleens, and brains were collected.
Mouse peripheral blood was obtained by cardiac puncture while spleen cells and CNS leucocytes were isolated as previously described [34] (link). Red blood cells were lysed with ACK lysing buffer (Gibco Cat # A1049201); cells were then stained with appropriate antibody cocktails against Alexa Fluor 647-CD11b (BioLegend Cat # 101220, RRID: AB_493546), Ly-6G-APC/Cy7 (Biolegend Cat # 127623, RRID: AB_10645331) and Ly-6C-PE/Cy7 (BioLegend Cat # 128017, RRID: AB_1732093) for 30 min at room temperature and analysed by flow cytometry. Cells were acquired in a FACS Canto II (Becton Dickinson). All analysis was carried out with FlowJo software (Tree Star).

Nowak W., Grendas L.N., Sanmarco L.M., Estecho I.G., Arena Á.R., Eberhardt N., Rodante D.E., Aoki M.P., Daray F.M., Carrera Silva E.A, & Errasti A.E. (2019). Pro-inflammatory monocyte profile in patients with major depressive disorder and suicide behaviour and how ketamine induces anti-inflammatory M2 macrophages by NMDAR and mTOR. EBioMedicine, 50, 290-305.

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Multicolor Flow Cytometry Analysis

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Single cell suspensions were prepared from spleens and pancreatic lymph nodes by disaggregation through 70 μm filters. The number of cells obtained from spleens and pancreatic lymph nodes were determined by flow cytometry by including AccuCount beads (Sphereotech, Lake Forest, IL) in the antibody-stained cell suspensions. The following antibodies were purchased from Biolegend (Nordic Biosite, Täby, Sweden): CD11b-Alexa700, Ly6G-APC-Cy7, F4/80-PE-Cy7, streptavidin Brilliant Violet 605. The following antibodies were purchased from BD Biosciences (San Jose, Ca, USA): CD19-PerCP-Cy5.5, Ly6C-biotin, SiglecF-PE. Prior to surface staining, cells were incubated with the 2.4G2 (anti-CD16/CD32) antibody to prevent unspecific binding. Cells were then stained with the above-mentioned antibodies in FACS buffer (PBS supplemented with 5% fetal calf serum and 0.05% NaN₃ (Sigma-Aldrich, St. Louis, MO). Fixable viability dye e-Fluor506 (eBioscience) was used for detecting and excluding dead cells from the analyses. Cells were analyzed using an LSRII flow cytometer (BD Biosciences) and FlowJo software (Tree Star, Ashland, OR).

Tahvili S., Törngren M., Holmberg D., Leanderson T, & Ivars F. (2018). Paquinimod prevents development of diabetes in the non-obese diabetic (NOD) mouse. PLoS ONE, 13(5), e0196598.

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Murine Model of Experimental Autoimmune Encephalomyelitis

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Myelin oligodendrocyte glycoprotein 35-55 (MOG_35-55) peptide (MEVGWYRSPFSRVVHLYRNGK) was generated by the Protein Core laboratory of the Blood Research Institute, BloodCenter of Wisconsin. The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780, CD25-Alexa Fluor 700, CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, CD11c-Biotin, B220-Biotin, CD8-Biotin, CD11b-biotin, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse B220-PE-Texas Red, IFN-γ-PE, anti-active caspase 3-FITC and anti-human Ki67-FITC were purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). Anti-mouse/rat Neuropilin-1-APC was obtained from R&D Systems (Minneapolis, MN). Monoclonal antibodies SMI-32 (anti-nonphosphorylated neurofilament-H) and SMI-99 (anti-myelin basic protein (MBP)) were purchased from Covance (Emeryville, CA). Strepavidin Alexa 405 and goat anti-mouse Alexa 546 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-Biotin microbeads were purchased from MiltenyiBiotec (Auburn, CA).

Ray A., Basu S., Miller N.M., Chan A.M, & Dittel B.N. (2014). An Increase in Tolerogenic Dendritic Cell and Natural T Regulatory Cell Numbers During EAE in Rras−/− Mice Results in Attenuated Disease. Journal of immunology (Baltimore, Md. : 1950), 192(11), 5109-5117.

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