Cells were stained in the dark on ice for 15 min with flow cytometry antibodies. Cells were then washed once with 1× PBS and resuspended in 1× PBS for sorting as described previously11 (link)–13 (link),73 (link). Antibodies used in this study were: PE anti-mouse CD45R/B220 (BD Biosciences, #553089, 1:100), PE anti-mouse TER-119 (Biolegend, #116207, 1:100), PE anti-O4 (R&D Systems, #FAB1326P, 1:100), PE anti-CD105 (eBioscience, #12–1051-82, 1:100), PE anti-CD140a (eBioscience, #12–1401-81, 1:100), PE anti-Ly-6G (Biolegend, #127608, 1:100), PerCP anti-Ly-6C (Biolegend, #128028, 1:100), APC anti-CD45 (eBioscience, #17–0451-83, 1:100), APC-Cy7 anti-CD11c (BD Biosciences, #561241, 1:100), and FITC anti-CD11b (eBioscience, #11–0112-85, 1:100). All cells were gated on the following parameters: CD105negCD140anegO4negTer119negLy-6GnegCD45Rneg. Astrocytes were subsequently gated on: CD11bnegCD45negLy-6CnegCD11cneg. Microglia were subsequently gated on: CD11bhighCD45lowLy-6Clow. Pro-inflammatory monocytes were subsequently gated on: CD11bhighCD45highLy-6Chigh. Compensation was performed on single-stained samples of cells and an unstained control. Cells were sorted on a FACS Aria IIu (BD Biosciences). For sorting of TdTomato+ astrocytes, cells were sorted according to TdTomato fluorescence judged against a wild-type control animal using a yellow-green laser on a FACS Aria IIu.
Anti cd11c apc cy7
Manufactured by BD
Sourced in United States
Anti-CD11c-APC-Cy7 is a fluorescently labeled antibody that binds to the CD11c surface antigen. It is used for the identification and analysis of cells expressing CD11c, such as dendritic cells, in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd45 apc, by Thermo Fisher Scientific (2 mentions)
Anti cd105 pe, by Thermo Fisher Scientific (2 mentions)
Pe anti cd140a, by Thermo Fisher Scientific (2 mentions)
Anti ly6g pe, by BioLegend (2 mentions)
Pe anti mouse cd45r b220, by BD (2 mentions)
Pe anti o4, by R&D Systems (2 mentions)
Percp anti ly 6c, by BioLegend (2 mentions)
Pe anti mouse ter 119, by BioLegend (2 mentions)
Fc block, by BD (2 mentions)
Anti cd11b fitc, by Thermo Fisher Scientific (1 mentions)
Facsaria iiu, by BD (1 mentions)
Zombie aqua fixable viability kit, by BioLegend (1 mentions)
Fc block, by Miltenyi Biotec (1 mentions)
Macsquant16, by Miltenyi Biotec (1 mentions)
Anti cd11c pe, by Miltenyi Biotec (1 mentions)
Anti epcam pe, by Miltenyi Biotec (1 mentions)
Cd86 apc, by Miltenyi Biotec (1 mentions)
Intracellular fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions)
Macsquant 10, by Miltenyi Biotec (1 mentions)
Celltrace violet, by Thermo Fisher Scientific (1 mentions)
7 protocols using anti cd11c apc cy7
1
Isolation and Identification of Murine Immune Cells
2
Flow Cytometric Analysis of Immune Cells
3
Comprehensive Immune Cell Analysis Protocol
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Cell suspensions were incubated for 15 min at 4°C with Fc Block (BD Biosciences) and then stained for 25 min at 4°C with the following fluorochrome-conjugated antibodies: anti-CD11c-APC-Cy7 (clone: HL3, BD Biosciences) or anti-CD11c-PE (clone REA754, Miltenyi Biotec), anti-MHC-II-VioBlue (clone: M5/114.15.2, Miltenyi Biotec), anti-CD11b-PerCP-Vio700 (clone: REA592, Miltenyi Biotec), anti-EpCAM-PE (clone: caa7-9G8, Miltenyi Biotec) or anti-EpCAM-PE-Vio770 (clone caa7-9G8, Miltenyi Biotec), anti-XCR1-APC-Vio700 (clone REA707, Miltenyi Biotec), anti-PD-L2-PE (clone MIH37, Miltenyi Biotec), CD86-APC (clone PO3.3, Miltenyi Biotec). Dead cells were excluded using Zombie Aqua Fixable Viability Kit (Biolegend). For the analysis of Fc receptor expression, cells were permeabilized using an intracellular fixation & permeabilization kit (eBioscience) and incubated for 25 min at 4°C with anti-CD16(FcγRIII)/CD32(FcγRII)-PE-Vio770 (instead of Fc Block, clone: 93, Miltenyi Biotec), anti-FcϵRIα-APC (clone: MAR-1, Miltenyi Biotec), anti-CD23-APC (clone: B3B4, Miltenyi Biotec), anti-CD64-PE-Vio770 (clone: REA286, Miltenyi Biotec). Cells were acquired on a MACSquant 10 or a MACSquant 16 flow cytometer (Miltenyi Biotec) and data were analyzed using FlowJo software using the gating strategies described in Figures S2 S3
4
Quantifying A20 Phagocytosis by DCs
5
Immune Profiling of Aortic Arch
6
Multiparametric Flow Cytometry Analysis
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Spleen and inguinal lymph node (LN) cells were harvested from immunized mice at the time of sacrifice. Joint tissues were minced and treated with collagenase (0.25 mg/ml), and joint cell populations were examined for surface markers using antibodies anti-CD4-AF700, anti-CD8a-PE-Cy7, anti-CD19-PE, anti-CD11b-Pacific Blue, anti-CD11c-APC-Cy7, anti-F4/80-APC, and anti-GR-1-FITC (BD Pharmingen, San Jose, CA, USA). For intracellular cytokine staining, cells were stimulated with Phorbol 12-myristate 13-acetate (25 ng/ml) and ionomycin (500 ng/ml) (Sigma-Aldrich) and treated with GolgiPlug (BD Pharmingen) for 4 hours. After cell surface staining with anti-CD3e-PE-Cy7 and anti-CD4-APC-Cy7, cells were permeabilized using the Cytofix/Cytoperm Plus kit (BD Pharmingen) and stained with anti-IFNγ-APC and anti-IL-17A-FITC. A BD LSRII cytometer was used for cytometry and data were analyzed using BD FACS Diva Software.
7
Flow Cytometric Analysis of Activated T Cells and Dendritic Cells
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Cells were washed with PBS (Life Technologies, Darmstadt, Germany) and dead cells were excluded by viability dye staining (eBioscienceTM Fixable Viability Dye eFluorTM 506) for 20 min on ice. Prior to the surface staining of CD8+ T cells or BMDCs, cells were washed twice with staining buffer (1% BSA, 0,02% NaN3 in PBS). To analyze activated CD8+ T cells, cells were stained for CD8 using anti-CD8-PB (eBioscience) for 30 min on ice. Thereafter, intracellular staining was performed, as described below. Alternatively, BMDCs were analyzed for their SIINFEKL/Kb-loading ability using anti-CD11c-PE, -I-A/I-E-PB, -Kb-PE/Cy7, and -SIINFEKL/H2-Kb-PE/Cy7 (all from eBioscience) or for their maturation phenotype using anti-CD11c-APC/Cy7, -CD86-APC, I-A/I-E-PE antibodies (all BD Pharmingen), or anti-CD40-PB antibody (Biolegend). Antibodies were added after blocking of Fc-receptors using CD16/32 antibodies (Fc-BlockTM, BD Biosciences). After incubation on ice for 30 min, cells were washed and fixed with 1% paraformaldehyde and stored at 8°C until flow cytometry was performed using BD FACS Canto II (BD Biosciences, Heidelberg, Germany).
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