Pe conjugated anti cd11c

Manufactured by Thermo Fisher Scientific

Sourced in United States

PE-conjugated anti-CD11c is a monoclonal antibody that binds to the CD11c antigen expressed on the surface of dendritic cells, macrophages, and some B cells. It is conjugated with the fluorescent dye Phycoerythrin (PE) for detection and analysis purposes.

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Close spelling variants of this product

Anti-CD11c (67 citations) CD11c-PE (37 citations) Anti-CD11c-PE (26 citations) PE-conjugated anti-mouse CD11c (4 citations) PE-conjugated CD11c (2 citations) PE-conjugated anti-CD11c Ab (2 citations) PE/Cy7-conjugated anti-CD11c (2 citations)

6 protocols using pe conjugated anti cd11c

Neutrophil Evaluation After TB Challenge

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The neutrophils from the vaccinated animals were evaluated 30 days after the challenge with M. tuberculosis. The splenocytes were obtained as described above. The cells were incubated for 30 min with APC-conjugated anti-GR1 (BD PharMingen), PercP-conjugated anti-CD14, PE-conjugated anti-CD11c and FITC-conjugated anti-CD11b antibodies (eBioscience). Then, the cells were washed with PBS and fixed with paraformaldehyde using Perm Fix (BD Cytofix). A total of 100,000 events were acquired using BD Biosciences FACSVerse^TM and analyzed with the FlowJo 9.0 Software. The neutrophils percentage was determined evaluating singlets that were GR-1⁺, CD11c^-, and CD14^-. The total number of neutrophils was obtained multiplying the percentage of neutrophils by the total number of splenocytes from each animal.

Trentini M.M., de Oliveira F.M., Kipnis A, & Junqueira-Kipnis A.P. (2016). The Role of Neutrophils in the Induction of Specific Th1 and Th17 during Vaccination against Tuberculosis. Frontiers in Microbiology, 7, 898.

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Isolation and Characterization of Immune Cells

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Cells from individual diaphragms were recovered following perfusion of blood from tissues, as described previously (22 (link)), and cultured ex vivo for 6 h with 250 ng/ml ionomycin (Sigma-Aldrich), 50 ng/ml PMA (Sigma-Aldrich), and 1 μg/ml brefeldin A (BD Pharmingen). After a 15 min incubation with Fc block (eBioscience) and 10% normal mouse serum, cells were incubated for 15 min with PE-Cy5-conjugated anti-CD4 (eBioscience). Samples were then treated with fixation/permeabilization buffer (eBioscience), and permeabilized cells were incubated for 1 h with PE-conjugated anti-IL-4 (eBioscience) or anti-IL-10 (eBioscience).
For dendritic cell phenotyping, cells from dLNs and diaphragms were incubated FITC-conjugated anti-CD11b, PE-conjugated anti-CD11c and Pacific blue-conjugated anti-MHCII. Data were acquired using a Gallios flow cytometer (Beckman Coulter) and analyzed with FlowJo software (Tree Star).

Huang L., Gebreselassie N.G., Gagliardo L.F., Ruyechan M.C., Lee N.A., Lee J.J, & Appleton J.A. (2014). Eosinophil-derived IL-10 supports chronic nematode infection. Journal of immunology (Baltimore, Md. : 1950), 193(8), 4178-4187.

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Phenotypic Analysis of Bone Marrow Dendritic Cells

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Bone marrow-derived dendritic cells were stained in FACS buffer (PBS containing 0.1% bovine serum albumin and 5 mM EDTA) using the following monoclonal antibodies in the indicated concentration for cell surface markers (all obtained from eBioscience): PE-conjugated anti-CD86 (2 μg/ml), FITC conjugated anti-CD-40 (5 μg/ml), PE-conjugated anti-CD11c (1 μg/ml), and APC-conjugated MHC-II (0.28 μg/ml). Cells were first incubated with Fc receptor block, 5 μg/ml (eBioscience) for 10 min to block any non-specific binding and subsequent staining steps were performed for 20 min at 4°C, followed by washing with FACS buffer. An isotype-matched control staining was done for each antibody to determine a specific background staining. Cells were acquired using a Cyan-ADP cytometer (Beckman Coulter, Woerden, The Netherlands) and analyzed with FlowJo software (FlowJo LLC, Ashland, OR, USA).

Wilbers R.H., Westerhof L.B., van de Velde J., Smant G., van Raaij D.R., Sonnenberg A.S., Bakker J, & Schots A. (2016). Physical Interaction of T Cells with Dendritic Cells Is Not Required for the Immunomodulatory Effects of the Edible Mushroom Agaricus subrufescens. Frontiers in Immunology, 7, 519.

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Dendritic Cell Phenotypic Analysis

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The BMM-derived DCs were generated as describe above and stained with FITC-conjugated anti-CD11c (eBioscience, Thermo Fisher Scientific, San Diego, CA, USA), PE-conjugated anti-CD11c (eBioscience), PE-conjugated anti-MHC class I (eBioscience), PE-Cy5.5-conjugated anti-CD86 (BioLegend, San Diego, CA, USA) or APC- conjugated anti-MHC class II (BioLegend) Ab. To evaluate the percentages of mature DCs in the draining lymph nodes and tumors, the cells expressed with CD11c⁺ and MHC class I^hi were gated as dendritic cells and then analyzed the co-expression of CD86⁺ and MHC class II^hi. Flow cytometric analyses were performed as previously described [37 (link),40 (link)].

Chen Y.L., Lin H.W., Sun N.Y., Yie J.C., Hung H.C., Chen C.A., Sun W.Z, & Cheng W.F. (2019). mTOR Inhibitors Can Enhance the Anti-Tumor Effects of DNA Vaccines through Modulating Dendritic Cell Function in the Tumor Microenvironment. Cancers, 11(5), 617.

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Isolation and Analysis of Alveolar Macrophages

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Isolated lung MNCs were stained with fluorescein isothiocyanate (FITC)-conjugated anti-F4/80 (Clone BM8, eBioscience, San Diego, CA, USA), phycoerythrin (PE)-conjugated anti-CD11c (Clone N418, eBioscience, San Diego, CA, USA) and allophycocyanin/cyanine7 (APC-Cy7)-conjugated anti-CD45 (Clone 104, Biolegend, San Diego, CA, USA). Subsequently, alveolar macrophages (CD45⁺ F4/80⁺ CD11c⁺) were sorted using a FACS Aria II flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). The purity of the separated cells was > 95%.

Chen Y., Hao X., Li M., Tian Z, & Cheng M. (2023). UGRP1-modulated MARCO+ alveolar macrophages contribute to age-related lung fibrosis. Immunity & Ageing : I & A, 20, 14.

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Psoriasis Induction and Analysis Protocol

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Aldara ® (IMQ) 5% cream (MEDA Pharmaceuticals, Vienna, Austria) was used for inducing psoriatic skin changes in the mice. 1% rapamycin (Merck Calbiochem, Darmstadt, Germany) ointment was made by mixing 50 mg petroleum jelly with 10 μl of the corresponding rapamycin stock, 50 mg/ml. P-mTOR S2448 #2976 and P-S6 S235/6 #2211 antibodies were from Cell Signaling Technology, Frankfurt, Germany. Keratin 6 antibody (Ks6. KA12) from Thermo Scientific, Darmstadt, Germany and caspase-14 antibody (NB100-56126) was from Novus Biologicals, Wiesbaden, Germany. NIMP-R14 Ly-6G/C (ab2557) was from Abcam, Cambridge, UK. Fluorescein isothiocyanate (FITC)conjugated anti-CD3, phycoerythrin (PE)-conjugated anti-CD4, PE-conjugated anti-CD11c, allophycocyanin (APC)-conjugated anti-CD11b, FITC-conjugated anti-F4/80, APC-conjugated anti-B220, PE-conjugated anti-langerin, APC-conjugated anti-EpCAM all were from eBioscience, Darmstadt, Germany, while peridinin chlorophyll protein complex (PerCP/Cy5)-conjugated anti-Siglec-H was from Biolegend, Koblenz, Germany and PerCP-conjugated anti-CD8 was purchased from BD Bioscience, Heidelberg Germany.

Bürger C., Shirsath N., Lang V., Diehl S., Kaufmann R., Weigert A., Han Y.Y., Ringel C, & Wolf P. (2017). Blocking mTOR Signalling with Rapamycin Ameliorates Imiquimod-induced Psoriasis in Mice. Acta dermato-venereologica, 97(9).

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