Flow cytometry analysis of tumor MNCs was performed using directly conjugated monoclonal mouse antibodies: anti-mouse F4/80 Pacific Blue (clone BM8), anti-mouse CD49b PerCP/Cy5.5 (clone DX5), anti-mouse CD11c FITC (clone N418), and anti-mouse CD3 APC/Cy7 (clone 17A2) (Biolegend, San Diego, CA, USA). Surface staining was performed according to the manufacturer’s instructions. For intracellular staining with IL-1β, the MNCs were fixed and permeabilized for 20 min at 4 °C with Cytoperm/Cytofix (BD Biosciences, Franklin Lakes, NJ, USA) and stained with anti-mouse IL-1β APC (Monoclonal Rat IgG2B Clone #166931) (R&D Systems, Minneapolis, MN, USA), antibodies in BD Perm/Wash solution according to the manufacturer’s instructions. Sample analysis was performed on a MACSQuant Analyzer (Miltenyi Biotec) and flow cytometry data were analyzed with FlowJo 8.7. software (FlowJo, LLC, New York, NY, USA).
Fitc anti mouse cd11c clone n418
Manufactured by BioLegend
Sourced in United States
FITC anti-mouse CD11c (clone N418) is a fluorescently labeled monoclonal antibody that binds to the CD11c antigen expressed on mouse dendritic cells. It can be used for the identification and analysis of dendritic cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa647 anti mouse cd45.1 clone a20, by BioLegend (2 mentions)
Percp cy5.5 anti mouse cd45 2 clone 104, by BioLegend (2 mentions)
Fcεr1α clone mar 1, by BioLegend (2 mentions)
Canto flow analyzer, by BD (2 mentions)
Pe cy7 anti mouse cd4 clone gk1.5, by BioLegend (2 mentions)
Facsaria cell sorter, by BD (2 mentions)
Cytofix cytoperm, by BD (1 mentions)
Anti mouse il 1β apc monoclonal rat igg2b clone 166931, by R&D Systems (1 mentions)
Macsquant analyzer, by Miltenyi Biotec (1 mentions)
P stat6 y641, by Cell Signaling Technology (1 mentions)
Apc anti mouse human cd11b clone m1 70, by BioLegend (1 mentions)
Pe rat anti mouse siglec f clone e50 2440, by BD (1 mentions)
Pe cy7 rat anti mouse cd45 clone 30 f11, by BD (1 mentions)
Apc anti mouse cd86 clone gl 1, by BioLegend (1 mentions)
Pe anti mouse cd80 clone 16 10a1, by BioLegend (1 mentions)
Tsg101, by Abcam (1 mentions)
6 protocols using fitc anti mouse cd11c clone n418
1
Tumor-infiltrating Immune Cell Analysis by Flow Cytometry
2
Immune Cell Profiling via Antibody Staining
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Anti-Mouse CD11c FITC (clone N418) was from BioLegend and used at a concentration of 1 μg/mL × 106 cells. Polyclonal rabbit anti-STAT1 (#9172), pSTAT1-Y701 (#9167), STAT6 (#9362), and pSTAT6-Y641 (#9361) were used at a dilution of 1/1000 and were from Cell Signaling. Polyclonal rabbit anti-iNOS (ab15323), anti-IFN-γ (EPR1108, ab133566), anti-MHC Class II (ab55152), and anti-CD36 (ab137320) were used at a dilution of 1/1000 and were from Abcam. Polyclonal goat anti-S100A8 (AF3059) and anti-S100A9 (AF2065) were used at a dilution of 1/1000 and were from R&D Systems. Polyclonal rabbit anti-MMP12 catalytic domain antibody and anti-hinge domain antibody (Triple Point Biologics) were used at a dilution of 1/1000. Anti-mouse myeloperoxidase (MPO) from Hycult (HM1051) were used at a dilution of 1/100 on tissue sections.
3
Phenotypic Characterization of Immune Cells
4
Multiparameter Flow Cytometry Profiling
5
Isolation and Characterization of Extracellular Vesicles from C57 Mice
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A total of 50 five-week-old male C57 mice (Animal Center of Southern Medical University, Guangzhou, China), weighing 25–30 g, were used in the present study. All mice were were acclimatized for 1 week before experiments at room temperature under a controlled 12/12 h light/dark cycle and received food and water ad libitum. Antibodies for western blot analysis were as follows: Rabbit monoclonal anti-mouse CD63 (cat. no. ab217345; Abcam), CD9 (cat. no. ab92726; Abcam), CD81 (cat. no. ab109201; Abcam) and TSG101 (cat. no. ab125011; Abcam). Antibodies for flow cytometry were as follows: FITC anti-mouse CD11c (clone N418; Biolegend), PE anti-mouse MHC-II (clone 10-3.6; Biolegend), PE anti-mouse CD80 (clone 16-10A1; Biolegend) and APC anti-mouse CD86 (clone GL-1; Biolegend).
All animal-related experiments were performed according to the guidelines of the Care and Use of Laboratory Animals (Ministry of Health, China, 1998) (14 (link)). The experiments were approved by the Animal Use Committee of Shenzhen Hospital, Southern Medical University.
All animal-related experiments were performed according to the guidelines of the Care and Use of Laboratory Animals (Ministry of Health, China, 1998) (14 (link)). The experiments were approved by the Animal Use Committee of Shenzhen Hospital, Southern Medical University.
6
Phenotypic Characterization of Immune Cells
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Single cell suspensions were obtained from MLNs, spleens, peritoneal lavage, and peripheral blood after bone marrow reconstitution and N. brasiliensis infection. Cells were blocked with unlabeled anti-mouse CD16/32 (clone 2.4G2) for 10min on ice, followed by incubation with fluorochrome labeled antibodies. Antibodies used were as follows Alexa647 anti-mouse CD45.1 (clone A20), PerCP-Cy5.5 anti-mouse CD45.2 (clone 104) (Biolegend), FITC lineage cocktail, CD127 (clone A7R34), ST2 (clone DIH9), CD117 (clone 2B8), and Fcεr1α (clone MAR-1) (Biolegend). Cells were analyzed on a BD Canto Flow analyzer; data was analyzed using FlowJo software (version7.6, TreeStar, Ashland, OR). For FACS, single cell suspensions of MLNs were stained with PE anti-mouse CD90.2 (clone 30.H12), APC anti-mouse/human B220 (clone RA3-6B2), PE-Cy7 anti-mouse CD4 (clone GK1.5), and FITC anti-mouse CD11c (clone N418) (Biolegend) and then sorted on a BD FACS Aria cell sorter (BD Biosciences, San Jose, CA).
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