Cd11c 3

Manufactured by Thermo Fisher Scientific

Sourced in United States

CD11c (3.9) is a monoclonal antibody that recognizes the CD11c antigen, which is expressed on the surface of various immune cells, including dendritic cells, monocytes, and macrophages. The antibody can be used for the identification and characterization of these cell populations in flow cytometry applications.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-CD11c Alexa700 (clone 3.9; (3 citations)

9 protocols using cd11c 3

Phenotypic Characterization of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monocyte purity, proper Mo-DC differentiation and the basal state of activation of both cells were assessed by flow cytometry using the following anti-human monoclonal antibodies: CD14 (M5E2, Biolegend, San Diego, CA, USA), CD1b (SN13, Biolegend), CD1d (51.1, Biolegend), CD11c (3.9, eBioscience), CD80 (2D10, Biolegend), HLA-DR (LN3, eBioscience).
iNKT and T cell determinations were performed in total PBMCs or in CD14⁻ fractions, using CD1d-PBS57 tetramers (NIH Tetramer Core Facility, Emory University, Atlanta, GA, USA) and the following anti-human monoclonal antibodies: CD3 (OKT3, eBioscience), CD4 (OKT4, Biolegend), and CD8 (RPA-T8, eBioscience). The purity of T cell clones VM-D5 and JS63 was assessed by using CD1d-PBS57 tetramers (NIH Tetramer Core Facility, Emory University, Atlanta, GA, USA) together with anti-human CD3 (OKT3, eBioscience) monoclonal antibody. The purity of T cell clones s33d, GG33A, and DS1C9b was assessed by using anti-human TCR Vβ13.1 (IMMV222), Vβ18 (BA62.6), and Vβ7.1 (ZOE), monoclonal antibodies from Immunotec (Immunotec Research Inc, Canada). Cells were acquired in a FACS Canto II (BD Biosciences, San Diego, CA, USA) using the BD FACSDiva™ software (BD Biosciences). Data analysis was performed with FlowJo® v10 (FlowJo LLC, Ashland, OR, USA).

Pereira C.S., Pérez-Cabezas B., Ribeiro H., Maia M.L., Cardoso M.T., Dias A.F., Azevedo O., Ferreira M.F., Garcia P., Rodrigues E., Castro-Chaves P., Martins E., Aguiar P., Pineda M., Amraoui Y., Fecarotta S., Leão-Teles E., Deng S., Savage P.B, & Macedo M.F. (2019). Lipid Antigen Presentation by CD1b and CD1d in Lysosomal Storage Disease Patients. Frontiers in Immunology, 10, 1264.

+ Open protocol

+ Expand

In Vitro Generation of Human Monocyte-Derived Dendritic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human MO-DC were generated in vitro as previously described (14 ). Briefly, CD14+ monocytes were magnetically isolated from PBMC (Miltenyi) and cultured in RPMI containing 10% FCS supplemented with GM-CSF (100ng/ml) and IL-4 (20ng/ml) (R&D Systems). Cultures were fed on day 4. On day 8, DC were seeded into poly(2-hydroxyethyl-methacrylate) (Sigma) coated 12-well culture plates (Corning Costar) at a density of 1 million cells/well and stimulated for 2 days with a cocktail containing IL-6 (10ng/ml), PGE₂ (0.1μM) along with IL-1β (10ng/ml), IL-36α, IL-36β or IL-36γ (100ng/ml) in 500μl complete RPMI. DC phenotype was analyzed by flow cytometry as described above using antibodies against CD86 (FUN-1, BD), HLA-DR (LN3, eBioscience), CD1a (HI149, eBioscience), CD11c (3.9, eBioscience), CD123 (6H6, Biolegend) and appropriate isotype control antibodies.

Foster A.M., Baliwag J., Chen C.S., Guzman A.M., Stoll S.W., Gudjonsson J.E., Ward N.L, & Johnston A. (2014). IL-36 promotes myeloid cell infiltration, activation and inflammatory activity in skin. Journal of immunology (Baltimore, Md. : 1950), 192(12), 6053-6061.

+ Open protocol

+ Expand

Immunocytochemistry of Thymic Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were cultured as above, but poly-L-lysine coated 96-well glass bottom plates (MatTek, Ashland, MA, USA) were used with 200 μl medium per well and 1–2 pieces of thymic tissue per well. Cells were fixed using 100 μl 1:1 4% paraformaldehyde/PBS for 5 min. Unspecific binding was blocked with serum-free protein block for 15 min (Dako, Copenhagen, Denmark). Cells were stained with Abs against EpCAM (HEA125), K8 (EP1628Y, Abcam, Cambridge, UK), K5 (D5/16 B4, Dako), HLA-DR, (G46-6, BD Pharmingen, Franklin Lakes, NJ, USA) or CD11c (3.9, eBioscience, San Diego, CA, USA) for 30 min at 37 °C. The wells were gently washed twice with PBS followed by incubation with alexa fluor-conjugated anti-mouse or anti-rabbit secondary antibodies for 30 min in 37 °C and finally washed in PBS and refilled with PBS. The stained cells were analyzed using a Zeiss lsm 700 at × 10x or × 25 magnification (Zeiss, Oberkochen, Germany).

Skogberg G., Lundberg V., Berglund M., Gudmundsdottir J., Telemo E., Lindgren S, & Ekwall O. (2015). Human thymic epithelial primary cells produce exosomes carrying tissue-restricted antigens. Immunology and Cell Biology, 93(8), 727-734.

+ Open protocol

+ Expand

Phenotypic Analysis of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For phenotypic analysis, cells were labelled with Abs recognizing CD11c (3.9), CD40 (5C3), CD45 (HI30), CD83 (HB15e), HLA-DR (LN3, all eBioscience), BDCA1/CD1c (AD5-8E7, Miltenyi Biotec), BDCA3/CD141 (AD5-14H12, Miltenyi Biotec), CD86 (2331, BD Horizon), CLEC9A (8F9, BioLegend), HBsAg (Acris), a lineage cocktail including CD3 (UCHT1, eBioscience), CD14 (61D3, eBioscience), CD19 (HIB19, eBioscience) and CD56 (MY31, BD Biosciences), and the live/dead marker Aqua (LifeTechnologies). Fluorescence was measured using a FACS Canto II (BD Biosciences).

van der Aa E., Buschow S.I., Biesta P.J., Janssen H.L, & Woltman A.M. (2016). The Effect of Chronic Hepatitis B Virus Infection on BDCA3+ Dendritic Cell Frequency and Function. PLoS ONE, 11(8), e0161235.

+ Open protocol

+ Expand

Phenotypic Profiling of Monocytes and DCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

1x10⁶ cryopreserved PBMC were thawed then washed twice in FACS buffer and surface stained using the following antibody cocktail: CD14 (M5E2, Biolegend), CD16 (3G8, Biolegend), CD11b (ICRF44, Biolegend), HLA-DR (L243, Biolegend), CD40 (5C3, BD Pharmingen), CD62L (DREG-56, BD Pharmingen), CD64 (10.1, Biolegend), CD163 (GHI/61, Biolegend), CXCR6 (K041E5, Biolegend), IFNAR (85228, R&D Systems), CD11c (3.9, ThermoFisher Scientific) and CD123 (6H6, Biolegend) for the detection of various monocytic and dendritic cell subsets. Samples were then acquired on the Attune NxT acoustic focusing cytometer (Life Technologies). Data were analyzed using FlowJo v10 (TreeStar, Ashland, OR USA).

Zulu M.Z., Sureshchandra S., Pinski A.N., Doratt B., Shen W, & Messaoudi I. (2021). Obesity Correlates With Pronounced Aberrant Innate Immune Responses in Hospitalized Aged COVID-19 Patients. Frontiers in Immunology, 12, 760288.

+ Open protocol

+ Expand

Flow Cytometric Profiling of PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Frozen PBMCs were thawed, washed in FACS buffer (2% FBS, 1mM EDTA in PBS), and counted on TC20 (Biorad) before surface staining using two independent flow panels. For the innate panel, the following antibodies were used: CD3 (SP34, BD Pharmingen) and CD20 (2H7, BioLegend) for the exclusion of T & B lymphocytes, respectively. We further stained for CD56 (HCD56, Biolegend), CD57 (HNK-1, BioLegend), KLRG1 (SA231A2, BioLegend) CD16 (3G8, BioLegend), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend) and CD86 (IT2.2, BioLegend). For the adaptive panel, the following antibodies were used: CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD45RA (HI100, TONBO Biosciences), CCR7 (G043H7, BD Biosciences), CD19 (HIB19, BioLegend), IgD (IA6–2, BioLegend), CD27 (M-T271, BioLegend), KLRG1 (SA231A2, BioLegend) and PD-1 (Eh12.2h7, BioLegend). Cells were stained with Ghost Dye viability dye (TONBO biosciences) for 30 minutes at 4C per manufacturer’s instructions, washed, surface stained with either innate or adaptive panels for 30 minutes at 4C. Samples were then washed and analyzed on Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA).

Sureshchandra S., Zulu M.Z., Doratt B., Jankeel A., Tifrea D., Edwards R., Rincon M., Marshall N.E, & Messaoudi I. (2021). Deep immune profiling of the maternal-fetal interface with mild SARS-CoV-2 infection. bioRxiv.

+ Open protocol

+ Expand

Immunophenotyping of Frozen PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Frozen PBMCs were thawed, washed in FACS buffer (2% FBS, 1 mM EDTA in PBS), and counted on TC20 (Biorad) before surface staining using two independent flow panels. For the innate panel, the following antibodies were used: CD3 (SP34, BD Pharmingen) and CD20 (2H7, BioLegend) for the exclusion of T & B lymphocytes, respectively. We further stained for CD56 (HCD56, Biolegend), CD57 (HNK-1, BioLegend), KLRG1 (SA231A2, BioLegend) CD16 (3G8, BioLegend), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend) and CD86 (IT2.2, BioLegend). For the adaptive panel, the following antibodies were used: CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD45RA (HI100, TONBO Biosciences), CCR7 (G043H7, Biolegend), CD19 (HIB19, BioLegend), IgD (IA6-2, BioLegend), CD27 (M-T271, BioLegend), KLRG1 (SA231A2, BioLegend) and PD-1 (Eh12.2h7, BioLegend). Cells were stained with Ghost Dye viability dye (TONBO biosciences) for 30 min at 4C per manufacturer’s instructions, washed, surface stained with either innate or adaptive panels for 30 min at 4C. Samples were then washed and analyzed on Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA).

Sureshchandra S., Zulu M.Z., Doratt B.M., Jankeel A., Tifrea D., Edwards R., Rincon M., Marshall N.E, & Messaoudi I. (2022). Single-cell RNA sequencing reveals immunological rewiring at the maternal-fetal interface following asymptomatic/mild SARS-CoV-2 infection. Cell Reports, 39(11), 110938.

+ Open protocol

+ Expand

Multiparametric Phenotyping of Decidual Leukocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

1-2 X 10⁶ fresh decidual leukocytes were washed with PBS and stained using the following cocktail of antibodies: CD45 (HI30, Biolegend), CD66b (G10F5, BioLegend), CD20 (2H7, BioLegend), CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD56 (HCD56, Biolegend), CD16 (3G8, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend), for 20 min in dark at 4C. Samples were washed twice in FACS buffer and resuspended in 400 uL. All samples were acquired with the Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA), immediately after addition of SYTOX Red Dead Cell Stain (1:1000). Data were analyzed using FlowJO (Ashland OR).

+ Open protocol

+ Expand

Comprehensive Immune Profiling of Decidual Leukocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

1–2 × 10⁶ fresh decidual leukocytes were washed with PBS and stained using the following cocktail of antibodies: CD45 (HI30, Biolegend), CD66b (G10F5, BioLegend), CD20 (2H7, BioLegend), CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD56 (HCD56, Biolegend), CD16 (3G8, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend), for 20 minutes in dark at 4C. Samples were washed twice in FACS buffer and resuspended in 400 uL. All samples were acquired with the Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA), immediately after addition of SYTOX Red Dead Cell Stain (1:1000). Data were analyzed using FlowJO (Ashland OR).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!