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Cd11c clone hl3

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CD11c (clone HL3) is a lab equipment product that functions as a cell surface marker. It is used to identify and study dendritic cells, which are a type of immune cell.

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Lab products found in correlation

Cd11b clone m1 70, by BD (4 mentions) Cd4 clone gk1, by BD (4 mentions) Cd4 clone rm4 5, by BD (3 mentions) Cd44 clone im7, by BD (3 mentions) Cd138 clone 281 2, by BD (3 mentions) Cd11b clone m1 70, by BioLegend (3 mentions) Cd4 clone gk1, by BioLegend (2 mentions) Fortessa, by BD (2 mentions) Cd69 clone h1.2f3, by BioLegend (2 mentions) B220 clone ra3 6b2, by BD (2 mentions) Cd23 clone b3b4, by BioLegend (2 mentions) Cd21 35 clone 7g6, by BD (2 mentions) F4 80 clone bm8, by Thermo Fisher Scientific (2 mentions) Facscalibur, by BD (2 mentions) Foxp3 intracellular staining kit, by Thermo Fisher Scientific (2 mentions) Cd45.2 clone 104, by BioLegend (2 mentions) Cd8 clone sk1, by BioLegend (2 mentions) Cd103 clone m290, by BioLegend (2 mentions) Cd172a clone p84, by BioLegend (2 mentions) Irf4 clone 3e4, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

CD11c (HL3; (41 citations) Anti-CD11c (clone HL3; (13 citations) CD11C APC (clone HL3, (6 citations) APC-anti-mouse-CD11c (clone HL3, (4 citations) Anti-CD11c–PE-Cy7 (clone HL3, (3 citations) APC-conjugated anti-CD11c (clone HL3), (3 citations) Anti-CD11c-PE (Clone HL3; (3 citations) CD11c FITC (clone HL3) (3 citations) Hamster anti-mouse CD11c (clone HL3, (3 citations) CD11c-APC-Cy7 (clone HL3) (2 citations)

21 protocols using cd11c clone hl3

1

Flow Cytometry Analysis of Immune Cell Subsets

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To detect surface proteins, mononuclear cells were incubated with Fc Block (Bio X Cell, 2.4G2) for 15 min and washed, followed by incubation with viability dye. The indicated antibodies were fluorescently conjugated against CD45 (clone 30-F11, BD Horizon, cat #563410), CD11b (clone M1/70, BD Biosciences, cat #563553), CD11c (clone HL3, BD Pharmingen, cat #553801), CD4 (clone GK1.5, BioLegend, cat #100428), CD8α (clone 53 – 6.7, BioLegend, cat #100734), CD19 (clone 6D5, BioLegend, cat #115541), and MHC Class II (clone M5/114.15.2, BioLegend, cat #107628). Samples were run on a BD Symphony (BD Biosciences) and analyzed using FlowJo software (Tree Star), as described (31 (link), 40 (link), 41 (link)).
Hong H., Wang Y., Menard M., Buckley J., Zhou L., Volpicelli-Daley L., Standaert D., Qin H, & Benveniste E. (2024). Suppression of the JAK/STAT Pathway Inhibits Neuroinflammation in the Line 61-PFF Mouse Model of Parkinson’s Disease. Research Square.
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2

Flow Cytometric Lineage Analysis of MPP

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Electronically sorted MPP were cultured on plates at 20,000 cells/ml, over OP9 or OP9Δ4 stromal cells at a 1:5 ratio, in OPTIMEM medium (Gibco) supplemented with 10% FCS, 1% antibiotics, 0.1% β-mercaptoethanol, SCF (1 ng/ml), Flt3L (10 ng/ml) (Immunotools), and IL-7 (8 ng/ml) (Peprotech, Neuilly-sur-Seine, France). After 7 days of incubation, cells were harvested, stained with appropriately labeled mAbs against CD4 (clone RM4-5), CD8 (clone 53-6.7), CD3 (clone 145-2C11), B220 (clone RA3-6B2), Gr1 (clone RB6-8C5), CD11c (clone HL3) all from BD Biosciences, NK1.1 (clone PK136, Sony) and CD11b (clone M1-70), ckit (CD117, clone 2B-8), Sca1(anti-Ly6A/E, clone D7) and PDCA-1 (clone eBio927) from eBioscience, and analyzed by flow cytometry for lineage determination.
Korniotis S., D’Aveni M., Hergalant S., Letscher H., Tejerina E., Gastineau P., Agbogan V.A., Gras C., Fouquet G., Rossignol J., Chèvre J.C., Cagnard N., Rubio M.T., Hermine O, & Zavala F. (2020). Mobilized Multipotent Hematopoietic Progenitors Stabilize and Expand Regulatory T Cells to Protect Against Autoimmune Encephalomyelitis. Frontiers in Immunology, 11, 607175.
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3

Multiparametric Flow Cytometry of Immune Cells

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Cell cultures were performed in complete culture medium consisting of RPMI-1640 supplemented with 10% heat-inactivated FCS, 2 mM L-glutamine (Sigma, St. Louis, MO), 50 μg/ml gentamicin (PAA, Linz, Austria), and 50 μM beta-mercaptoethanol (Sigma).
Phenotypical analyses were performed by flow cytometry with mAb against CD4 (clone RM4-5), CD45.1 (clone A20), MHC class II (anti-I-A/I-Ediverse, clone 2G9), CD11c (clone HL3), CD8α (clone Ly-2), CD45 (clone 30-F11), CD103 (clone M290), EpCAM/CD326 (clone G8.8), CD19 (clone 1D3), NK1.1 (clone PK136), IL-12p40 (clone C15.6) (all from BD-Pharmingen, San Diego, CA), CD127 (clone A7R34, Biolegend, San Diego, CA), CD70 (clone FR70, Biolegend), and Langerin/CD207 mAb (clone 929F3; Dendritics, Lyon, France). When possible, viable cells were determined by exclusion of 7-AAD-positive dead cells (BD-Pharmingen). IL-12p40 and IL-10 stainings were performed on total lymph node cell suspension (106 cells/ml) incubated for 3 h in 1 μg/ml Brefeldin A (Golgiplug, BD-Pharmingen). To stain for Langerin or intracellular cytokines, permeabilization was performed with Cytofix/perm kit (BD-Pharmingen).
Flacher V., Tripp C.H., Mairhofer D.G., Steinman R.M., Stoitzner P., Idoyaga J, & Romani N. (2014). Murine Langerin+ dermal dendritic cells prime CD8+ T cells while Langerhans cells induce cross-tolerance. EMBO Molecular Medicine, 6(9), 1191-1204.
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4

Multiparametric Flow Cytometry Analysis

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The frequency of immunological cellular subpopulations in pleural cells was analyzed by flow cytometry. Briefly, cells were stained for 30 min at 4°C with different combinations of the following fluorochrome-conjugated mAb: GR-1 (Clone RB6-8C5), F4/80 (Clone BM8), Ly6C (Clone HK1.4), CD3 (Clone 145-2c11), and CD4 (Clone GK1.5) (BioLegend), and CD11c (Clone HL3) (BD Bioscience), and iNOS (Clone CXNFT) (Cell Signaling technology, eBioscience). After antibody incubation, cells were washed with PBA (phosphate buffered saline containing 0.1% Sodium Azide and 0.2% Albumin bovine). Data were collected using a FACs CyAn flow cytometer (Beckman Coulter, Inc.) and analyzed with FlowJo (Tree Star) software. 100,000 events were acquired per sample.
Chavez-Galan L., Vesin D., Uysal H., Blaser G., Benkhoucha M., Ryffel B., Quesniaux V.F, & Garcia I. (2017). Transmembrane Tumor Necrosis Factor Controls Myeloid-Derived Suppressor Cell Activity via TNF Receptor 2 and Protects from Excessive Inflammation during BCG-Induced Pleurisy. Frontiers in Immunology, 8, 999.
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5

Comprehensive Immune Cell Analysis

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Spleens and cLNs were harvested and dissociated by mechanical dispersion. Following RBC lysis with ACK Lysis Buffer (Lonza), cells were incubated with Fc block (CD16/32, clone 2.4G2, BD Biosciences) and treated with antibodies directed against the following as indicated: B220 (clone RA3-6B2, BD Biosciences), CD23 (clone B3B4, Biolegend), CD21/35 (clone 7G6, BD Biosciences), T-bet (clone 4B10, BD Biosciences), CD11c (clone HL3, BD Biosciences), CD4 (clone GK1.5, BD Biosciences), CD8α (clone 53-6.7, BD Biosciences), CD44 (clone IM7, BD Biosciences), CD62L (clone MEL-14, BD Biosciences), CD138 (clone 281-2, BD Biosciences), CD69 (clone H1.2F3, Biolegend), and TLR7 (clone A94B10, BD Biosciences). Data were acquired using a BD Biosciences Fortessa and analyzed with FlowJo software (BD Biosciences).
Punnanitinont A., Kasperek E.M., Kiripolsky J., Zhu C., Miecznikowski J.C, & Kramer J.M. (2022). TLR7 agonism accelerates disease in a mouse model of primary Sjögren’s syndrome and drives expansion of T-bet+ B cells. Frontiers in Immunology, 13, 1034336.
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6

Multiparameter Flow Cytometry Isolation

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Flow cytometry was performed as previously described [9 (link)]. Spleen and cervical lymph nodes (cLNs) were mechanically dissociated and treated with RBC lysis buffer. Cells were incubated with Fc block (Cd16/32, clone 2.4G2, BD Biosciences) and were treated with the following antibodies as indicated: B220 (clone RA3-6B2, BD Biosciences), CD23 (clone B3B4, BioLegend), CD21/35 (clone 7G6, BD Biosciences), CD4 (clone GK1.5, BD Biosciences), CD69 (clone H1.2F3, BioLegend), CD86 (Clone GL1, BD Biosciences), CD11c (clone HL3, BD Biosciences), CD11b (clone M1/70, BD Biosciences), and CD138 (Clone 281-2, BD Biosciences). Data were acquired using a Fortessa (BD Biosciences) and analyzed using FlowJo software (BD Biosciences).
Sort-purification was performed using a FACSAria (BD Biosciences). Splenocytes were incubated with antibodies directed against B220, CD4, CD11b and CD11c as described above. B cells (B220+), T cells (CD4+) or monocytes (CD11b+, CD11c+, and CD11b/CD11c double positive) (1 × 105 cells) were sorted directly into lysis buffer for RNA isolation (RNeasy Plus Mini Kit, Qiagen). For stimulation experiments, B220+ cells were sorted into media and cultured overnight with either culture media alone or culture medium containing either LPS or anti-IgM/IgG + IL-4, as described above.
Kiripolsky J., Kasperek E.M., Zhu C., Li Q.Z., Wang J., Yu G, & Kramer J.M. (2021). Tissue-specific activation of Myd88-dependent pathways governs disease severity in primary Sjögren’s syndrome. Journal of autoimmunity, 118, 102608.
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7

Multicolor Flow Cytometry Staining

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For multicolor staining, fluorocrome-conjugated Abs (BD-Pharmingen, La Jolla, CA) against lineage markers were used in various combinations. Briefly, Fc receptors on cells were blocked with an anti-CD16/32 Ab for 30 min at 4°C and washed. Cells were then stained for surface markers for 30 min at 4°C, washed twice and analyzed by flow cytometry with a BD FACS Canto™ II cytometer (BD Biosciences, San José, CA, USA). For flow cytometry cell sorting, splenocytes from IL-12+IL-18-cDNA treated mice were stained with fluorocrome-labeled CD4 (clone RM4-5), CD8 (clone 53-6.7), CD11b (clone M1/70) and CD11c (clone HL3) antibodies (BD-Pharmingen, La Jolla, CA) and sorted to a purity of 97–99% using a MoFlo sorter (Dako Cytomation).
Barrios B., Baez N.S., Reynolds D., Iribarren P., Cejas H., Young H.A, & Rodriguez-Galan M.C. (2014). Abrogation of TNFα Production during Cancer Immunotherapy Is Crucial for Suppressing Side Effects Due to the Systemic Expression of IL-12. PLoS ONE, 9(2), e90116.
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8

Comprehensive Immune Cell Phenotyping

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Cell suspensions were stained in ice cold PBS with or without 2 % FBS on ice. The following antibodies were used for staining mouse cells: CD45 (clone 30-F11, Biolegend), Ly6G (clone 1A8, BD Biosciences), CD11c (clone HL3, BD Biosciences), CD11b (clone M1/70, Biolegend), Siglec F (clone E50–2440, BD Biosciences), CD3e (clone 145–2C11, eBioscience), CD4 (clone GK1.5, Biolegend), IFN © (clone XMG1.2, Biolegend), IL-17 (clone eBio17B7, eBioscience), IL-5 (clone TRFK5, Biolegend), IL-13 (clone eBio13A, eBioscience), CD25 (clone PC61.5, eBioscience), CD44, CD62L (clone MEL-14, BD Biosciences), MHC class II I-A/I-E (clone M5/114.15.2, eBioscience), DC-SIGN/CD209 (R&D systems), CD32 (clone D34–485, eBioscience), TLR4/CD284 (clone SA15–21, Biolegend), Annexin V (Biolegend). Neutrophils were defined as CD45+CD11b+Ly6G+DNA+ and cytoplasts were defined at CD45+CD11b+Ly6G+DNA−. Naïve T lymphocytes were defined as CD4+CD25−CD44lowCD62Lhi. Lung Dendritic cells were defined as CD45+CD11c+ MHCII+autofluorescencelow. The following antibodies were used to stain human cells: anti-CD45 PE-Cy7 (HI30), anti-CD66b (G10F5), anti-CD16 APC-Cy7 (3G8) all from Biolegend. Vybrant DyeCycle Ruby (Life Technologies) was used to stain intracellular DNA. Data were acquired on BD Canto II or BD LSR Fortessa and analyzed using FlowJo v10. For cell sorting, FACS Aria was used.
Krishnamoorthy N., Douda D.N., Brüggemann T.R., Ricklefs I., Duvall M.G., Abdulnour R.E., Martinod K., Tavares L., Wang X., Cernadas M., Israel E., Mauger D.T., Bleecker E.R., Castro M., Erzurum S.C., Gaston B.M., Jarjour N.N., Wenzel S., Dunican E., Fahy J.V., Irimia D., Wagner D.D, & Levy B.D. (2018). Neutrophil cytoplasts induce Th17 differentiation and skew inflammation toward neutrophilia in severe asthma. Science immunology, 3(26), eaao4747.
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9

Skin Histology and Immunohistochemistry

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Dorsal skin was collected and processed as described previously for histology and immunohistochemistry44 (link). Formalin-fixed, paraffin-embedded skin was sectioned and stained with haematoxylin and eosin (H&E) using standard techniques as described44 (link). Fresh frozen skin was sectioned and stained, as previously published44 (link), 49 (link), using antibodies targeting the following proteins: CD4 (clone RM4-5, Cat.#550280), CD8a (clone 53-6.7, Cat.#550281), CD11c (clone HL3, Cat.#550283; all BD Pharmingen; San Jose, CA) and F4/80 (clone BM8, Cat.#14-4801, eBioscience; San Diego, CA).
Epidermal thickness (acanthosis) measurements and immune cell quantification in dorsal skin sections was completed using microscopic images collected from the stained sections using interactive image analyses approaches as described previously44 (link).
Arbiser J.L., Nowak R., Michaels K., Skabytska Y., Biedermann T., Lewis M.J., Bonner M.Y., Rao S., Gilbert L.C., Yusuf N., Karlsson I., Fritz Y, & Ward N.L. (2017). Evidence for biochemical barrier restoration: Topical solenopsin analogs improve inflammation and acanthosis in the KC-Tie2 mouse model of psoriasis. Scientific Reports, 7, 11198.
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10

Bone Marrow-Derived Dendritic Cell Characterization

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BM-DCs derived from WT, TLR2KO and MyD88KO mice were cultured and assessed as previously described [29 (link)]. Briefly, mouse bone marrow cells were cultured at a density of 2 × 106 cells in Petri dishes containing 10 ml of complete RPMI-1640 supplemented with 200 unit/ml (20 ng/ml) recombinant mouse GM-CSF (PeproTech, Rocky Hill, NJ, USA). An additional 10 ml of complete RPMI containing 20 ng/ml GM-CSF was added on day 3. The cells were collected from each dish and counted on day 6. BM-DCs (1 × 106 cells/ml) were stimulated with the indicated concentrations of rOVA, rlipo-OVA, LPS, Pam3CSK4 or medium for 18 h. Cell surface markers [CD11c (clone HL3, BD Biosciences, San Jose, CA, USA), CD40 (clone 3/23, BD Biosciences) and CD80 (clone B7-1, eBioscience, San Diego, CA, USA)] of BM-DCs were analyzed using the FACSCalibur (BD Biosciences). The production of cytokines by BM-DCs (TNF-α and IL-12p40) was determined using ELISA kits (eBioscience).
Wu C.C., Liu S.J., Chen H.W., Shen K.Y, & Leng C.H. (2016). A Toll-like receptor 2 agonist-fused antigen enhanced antitumor immunity by increasing antigen presentation and the CD8 memory T cells population. Oncotarget, 7(21), 30804-30819.
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