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Cd11c pe cy7

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CD11c-PE-Cy7 is a fluorescently labeled monoclonal antibody that binds to the CD11c antigen expressed on the surface of dendritic cells and certain other myeloid cells. This antibody can be used in flow cytometry applications to identify and quantify CD11c-positive cell populations.

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Lab products found in correlation

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Close spelling variants of this product

CD11c-PE (57 citations) Anti-CD11c-PE-Cy7 (15 citations) Anti-CD11c-PE-Cy7 (HL3, (6 citations) PE-Cy7-conjugated anti-CD11c (6 citations) Anti-CD11c–PE-Cy7 (clone HL3, (3 citations) PE-Cy7-conjugated CD11c (3 citations) PE-Cy7-conjugated anti-mouse CD11c (3 citations) CD11c PE‐Cy7 (HL3, (2 citations) CD11c-PE-Cy7 (clone HL3, (2 citations) Anti-CD11c-PE-cy7 antibody (Ab) (2 citations)

65 protocols using cd11c pe cy7

1

Splenic Immune Cell Phenotyping

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Spleens were prepared and stained as described [21] (link) using the following surface antibodies for cell enumeration: biotinylated Siglec H (Hycult Biotech) and streptavidin-APC (BD Biosciences), CD11c PE-Cy7 (BD Biosciences), CD11b PerCP-Cy5.5 (Biolegend), CD19 APC-Cy7 (BD Biosciences), CD3e Pacific Blue (Biolegend), Ter-119 FITC (Biolegend). For intracellular cytokine staining the following antibodies were used: biotinylated Siglec H (streptavidin-APC), CD11c PE-Cy7, CD11b PerCP-Cy5.5, CD19 APC-Cy7, TNF Alexa 700 (BD Biosciences), IL-12p40 PE (BD Biosciences), IFNalpha FITC (Antigenix).
Snyder J.M., Treuting P.M., Nagy L., Yam C., Yi J., Brasfield A., Nguyen L.P, & Hajjar A.M. (2014). Humanized TLR7/8 Expression Drives Proliferative Multisystemic Histiocytosis in C57BL/6 Mice. PLoS ONE, 9(9), e107257.
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2

Comprehensive Immune Cell Profiling

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The following antibodies were used: CD4-V500, CD11c-PE-Cy7, CD19-V450, CD19-APC-Cy7, CD69-PE-Cy7, IgM-PerCP-Cy5.5, CD45.2-V500, CD45.2-APC (BD Biosciences), IgD-PE, CD45.1-FITC, CD11b-PerCP-Cy5.5, CD21-PB, CD23-PE-Cy7, AA4.1-APC (eBiosciences), and CD86-PB, CD45.1-PB (BioLegend). Following red blood cell lysis, Fc receptor blockade, and staining, cells were processed on a BD FACSVerse or LSR II flow cytometer and analyzed using FlowJo v9.6.4 (Treestar).
Mills R.E., Lam V.C., Tan A., Cresalia N., Oksenberg N., Zikherman J., Anderson M., Weiss A, & Hermiston M.L. (2015). Unbiased modifier screen reveals that signal strength determines the regulatory role murine TLR9 plays in autoantibody production. Journal of immunology (Baltimore, Md. : 1950), 194(8), 3675-3686.
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3

Tolerogenic Dendritic Cell-Induced T Cell Proliferation

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DCs were loaded with 1 mM liposomes (empty or loaded with insulin peptides) during 2 hours in the presence of insulin (20μg/ml, Sigma). DCs were cultured in basal conditions or matured with LPS (100 ng/ml, Sigma) for additional 24 hours to determine tolerogenic function stability. T cells were obtained after mechanical disruption of spleen and purified by negative selection using antibodies to CD19-PE, CD16/32-PE, CD11c-PECy7 (BD Biosciences), CD11b-PE (ImmunoTools GmbH, Friesoythe, Germany), and sorted (FACSAria II, BD Biosciences) as previously described [8 (link)]. 104 DCs were then cocultured with 105 T lymphocytes (1:10 ratio). As a control, T lymphocytes (105) were cultured in basal conditions. After 5 days, cells were pulsed with 1 μCi of (3H)-thymidine (Perkin Elmer, Waltham, MA) for an additional 16 hours. Cells were harvested (Harvester 96, Tomtec Inc., Hamden, CT) and analyzed using a scintillation counter (1450 Microbeta, TriluxWallac, Turku, Finland). T cell proliferation was expressed as counts per minute (c.p.m). Cytokine production was assessed using The Mouse Th1/Th2/Th17 kit (CBA system; BD Biosciences) in supernatants from proliferation assays. Data were analyzed using CBA software. The production of TGF-β was determined using Human/Mouse TGF-β1 Ready-SET-Go! (eBioscience).
Pujol-Autonell I., Serracant-Prat A., Cano-Sarabia M., Ampudia R.M., Rodriguez-Fernandez S., Sanchez A., Izquierdo C., Stratmann T., Puig-Domingo M., Maspoch D., Verdaguer J, & Vives-Pi M. (2015). Use of Autoantigen-Loaded Phosphatidylserine-Liposomes to Arrest Autoimmunity in Type 1 Diabetes. PLoS ONE, 10(6), e0127057.
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4

Multiparametric Phenotyping of Cervical Immune Cells

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Human cervical tissue was obtained from two sets of five healthy women (age range 42–47 years old) undergoing hysterectomy for benign indication at HUGTP. After confirmation of healthy tissue status by the Pathological Anatomy Service, a piece from ecto and endocervix separated by anatomical localization was delivered to the laboratory in refrigerated RPMI 1640 medium (Cellgro, Manassas, VA) containing 10% FBS (Lonza, Basel, Switzerland), 500U/mL penicillin, 500μg/mL streptomycin, 5μg/mL fungizone and 1μg/mL gentamycin (Life Technologies). Tissue was processed within the next 12 h after surgery, and 8-mm3 block-dissection was performed as described [26 (link)]. Tissue digestion of 5–9 pieces of ecto or endocervix with collagenase IV (Invitrogen) was immediately executed as described [26 (link)]. Tissue blocks were then dissociated manually with a disposable pellet pestle and filtered through a 70μm-cell strainer (BD Biosciences). After centrifugation, pellet was suspended in staining buffer (1% mouse serum, 1% goat serum in PBS) and stained with different combinations of CD3-eFluor 605 (eBiosciences), CD14-V450, CD11c-PE-Cy7, CD8-V500, HLA-DR-PerCP-Cy5.5, CD69-Horizon PE-CF594 (BD Biosciences), NK1.1-PE, γδTCR-FITC, Vα7.2-APC-H7 (Miltenyi Biotec), CD103-FITC and Vα24-APC (BioLegend). Data were acquired and analyzed as described for blood.
Qualai J., Li L.X., Cantero J., Tarrats A., Fernández M.A., Sumoy L., Rodolosse A., McSorley S.J, & Genescà M. (2016). Expression of CD11c Is Associated with Unconventional Activated T Cell Subsets with High Migratory Potential. PLoS ONE, 11(4), e0154253.
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5

Comprehensive Immune Profiling by Flow Cytometry

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1 × 106 splenocytes were taken for antibody staining and downstream analysis. Specific monoclonal antibodies were purchased from eBioscience (Thermofisher Scientific, Waltham, MA, USA) or BD Biosciences (San Jose, CA, USA): CD4 APC-Cy7 (RM4-5), CD8 BV510 (53-6.7), CD11b BV510 (M170), CD11c PECy7 (HL3), CD19 PerCP-Cy5.5 (ID3), CD69 BV421 (H1.2F3), CD279 PE (PD1; J43), Gr-1 Alexa700 (Ly6G/Ly6C; RB6-8C5), MHC-II FITC (I-A/I-E; 2G9) and CD16/32 (2.4G2). H-2Db restricted LCMV tetramer staining and peptide restimulation were performed as previously described [20 (link)]. All flow cytometry data were collected on a Fortessa X20 or Fortessa1 (BD Biosciences) and analyzed using FlowJo software (version 10, Flowjo LLC, Ashland, OR, USA).
Preston S.P., Allison C.C., Schaefer J., Clow W., Bader S.M., Collard S., Forsyth W.O., Clark M.P., Garnham A.L., Li-Wai-Suen C.S., Peiris T., Teale J., Mackiewicz L., Davidson S., Doerflinger M, & Pellegrini M. (2023). A necroptosis-independent function of RIPK3 promotes immune dysfunction and prevents control of chronic LCMV infection. Cell Death & Disease, 14(2), 123.
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6

Characterization of Bone Marrow-Derived Dendritic Cells

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On day 5–6 of BM-DC culture, cells were gently removed and stained with the following antibodies (or appropriate isotype controls): CD16/CD32 Fc block (BD Biosciences, San Jose, CA), CD11c-APC (BD Biosciences), CD11c-PE-Cy7 (BD Biosciences), CD11b-PE (BD Biosciences), I-Ad-FITC (BD Biosciences), CD86-APC (BioLegend, San Diego, CA) or CD80-PE (BD Biosciences). Mean Fluorescence Intensity (MFI) of the above markers was determined on CD11c+ gated cells. On day 7, Leukocyte Activation Cocktail (PMA/Ionomycin/Golgi-plug; BD Biosciences) was added to culture media for the final 4 hours of culture. Cells were gently removed and stained with CD16/CD32 Fc block followed by CD11c-PE-Cy7. Cells were permeabilized and fixed (eBioscience, San Diego, CA) and then stained with anti-IL-12-PE (BD Biosciences) or anti-IL-6-PE (BD Biosciences).
Looney B.M., Chernatynskaya A.V., Clare-Salzler M.J, & Xia C.Q. (2014). Characterization of Bone Marrow-Derived Dendritic Cells Developed in Serum-Free Media and their Ability to Prevent Type 1 Diabetes in Nonobese Diabetic Mice. Journal of blood disorders & transfusion, 5(4), 206.
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7

Quantifying Immune Cells in SARS-CoV-2 Infected Lungs

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Homogenized lungs suspensions from lungs of SARS-CoV-2 infected mice were rinsed and resuspended in PBS prior to staining with the following anti-mouse antibody cocktail for 30 min on ice: CD45-BV510 (BD; BDB563891), CD11b-FITC (BD; BDB557396), CD11C-Pe/Cy7 (BD; BDB558079), Ly6C-PerCP/Cy5.5 (BD; BDB560525), Ly6G-Alexa Fluor 700 (BD; BDB561236). Stained samples were then rinsed with PBS and resuspended in 4% PFA and allowed to fix for 72 h in accordance with biosafety requirements before analyzing using a BD LSRFortessa™ X-20 Cell Analyzer. Flow data was quantified using FlowJo (Tree Star). Bioinformatic analysis was performed using GraphPad Prism 5.0 (GraphPad).
Badeti S., Jiang Q., Naghizadeh A., Tseng H.C., Bushkin Y., Marras S.A., Nisa A., Tyagi S., Chen F., Romanienko P., Yehia G., Evans D., Lopez-Gonzalez M., Alland D., Russo R., Gause W., Shi L, & Liu D. (2022). Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection. Cell & Bioscience, 12, 88.
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8

Immune Cell Analysis in Musculoskeletal Tissue

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To analyze immune cells in the musculoskeletal tissues at 3 and 7 dpi, ipsilateral feet were perfused extensively, skinned, disjointed from the tibia, and digested in RPM11640 medium supplemented with 10% FBS, 15 mM HEPES, 0.5 mg/ml of collagenase (Sigma, C0130) and 10 ug/ml of DNase I (Sigma, D5025) for 1 h at 37°C. Digested tissue was passed through a 70 μm strainer, and cells were separated by centrifugation and washed with PBS supplemented with 2% FBS and 2 mM EDTA. Cells were stained with a Fixable Viability Dye eFluor 506 (eBioscience, 65-0866-14) and incubated with the following antibodies for 1 h at 4°C: CD16/32 (Biolegend, 101301, 1:200) CD11b PE-Dazzle 594 (BioLegend, 101256, 1:200), Ly6G PerCP-Cy5.5 (Biolegend, 127616, 1:400), Ly6C Pacific Blue (BioLegend, 128014, 1:200), F4/80 APC (Thermo Fisher, 17-4801-82, 1:200), CD11c PE-Cy7 (BD Biosciences, 558079, 1:200), I-A/I-E (MHC class II) Alexa Fluor 700 (BioLegend, 107622, 1:200), NK-1.1 PE (BioLegend, 108708, 1:200), CD3 BUV737 (BD Biosciences, 564380, 1:200), CD8 PerCP-Cy5.5 (BioLegend, 100734, 1:200), CD4 BV786 (BioLegend, 100552, 1:200), B220 BV711 (BioLegend, 103255, 1:200). Cells were analyzed using a BD X20 Fortessa flow cytometer, and all data were processed using FlowJo software (FlowJo, LLC).
Zhang R., Earnest J.T., Kim A.S., Winkler E.S., Desai P., Adams L.J., Hu G., Bullock C., Gold B., Cherry S, & Diamond M.S. (2019). Expression of the Mxra8 Receptor Promotes Alphavirus Infection and Pathogenesis in Mice and Drosophila. Cell reports, 28(10), 2647-2658.e5.
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9

Flow Cytometric Profiling of Adipose Tissue

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Dissected adipose tissue was cut into small pieces and suspended in RPMI (Sigma, USA) with 20% heat-inactivated FCS (Lonza, Switzerland) before homogenization for 1 hour in a shaker-incubator at 37 °C with 2 mg/mL collagenase II (C6885, Sigma, USA). Homogenized adipose tissue was processed as previously described55 (link) using FACS buffer (PBS with 1% heat-inactivated FCS and 0.1% NaN3) without fixation and permeabilisation. Cells were recorded on a LSRII (BD Biosciences, USA) flow cytometer, and data further analysed using Flowjo software (V10.0.7, Treestar). Gating strategies are shown in Fig. S6. The following antibodies for surface staining were used: CD45/PerCP (BioLegend, 30-F11), Siglec-F/PE (BD, E50-2440), CD11b/V500 (BD, M1/70), F4/80/APC (BioLegend, BM8), CD11c/APC-Cy7 (BD, HL3), CD45/AF647 (BD, 30-F11), CD4/AF488 (eBioscience, GK1.5), CD8a/FITC (BD, 53–6.7), CD11b/FITC (eBioscience, M1/70), CD49b/FITC (eBioscience, HMa2), F4/80/FITC (eBioscience, BM8), NK1.1/FITC (BD, PK136), FcεR1/PerCP-eF710 (eBioscience, MAR-1), CD19/PerCP-Cy5.5 (eBioscience, 1D3), CD11c/PE-cy7 (BD, HL3), ST2-biotin (MD biosciences, 101001B), Streptavidin-PE (eBioscience).
Banhos Danneskiold-Samsøe N., Sonne S.B., Larsen J.M., Hansen A.N., Fjære E., Isidor M.S., Petersen S., Henningsen J., Severi I., Sartini L., Schober Y., Wolf J., Nockher W.A., Wolfrum C., Cinti S., Sina C., Hansen J.B., Madsen L., Brix S, & Kristiansen K. (2019). Overexpression of cyclooxygenase-2 in adipocytes reduces fat accumulation in inguinal white adipose tissue and hepatic steatosis in high-fat fed mice. Scientific Reports, 9, 8979.
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10

Phenotypic Analysis of T Cells

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For phenotypic analysis of T cells, the PPD and peptides stimulated lungs and spleen/LNs cells were analysed by flow cytometry. Lymphocytes culture were harvested and stained with fluorochrome tagged anti-CD4-PE, CD8-APCCy7, CD62L-FITC, CD44-PerCPCy5.5, CD11c-PECy7, F4/80-APC, CD86-PE, CD80-FITC, CD40-PECy5, and MHC-II-PerCPCy5.5abs (BD Biosciences, San Jose, CA). Briefly, lymphocytes were harvested in tubes and washed with FACS buffer (PBS + 2%FCS). Cells were Fc blocked using anti-mouse CD16/CD32 Ab. Later, stained with fluorochrome-labelled Abs. After staining, cells were fixed by using 1% paraformaldehyde in FACS buffer. Cells were acquired in BD-FACS Aria III and BD-FACS Accuri (BD, Franklin Lakes, NJ). The analysis was performed using BD-FACS DIVA, BD-C6, and Flowjo software (BD, Franklin Lakes, NJ).
Maurya S.K., Aqdas M., Das D.K., Singh S., Nadeem S., Kaur G, & Agrewala J.N. (2020). A multiple T cell epitope comprising DNA vaccine boosts the protective efficacy of Bacillus Calmette–Guérin (BCG) against Mycobacterium tuberculosis. BMC Infectious Diseases, 20, 677.
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