Anti mouse cd86 pe

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-mouse CD86-PE is a flow cytometry reagent used to detect the expression of the CD86 cell surface marker in mouse cells. CD86 is a co-stimulatory molecule involved in T-cell activation. This reagent is conjugated with the fluorescent dye phycoerythrin (PE) to facilitate detection and quantification of CD86-positive cells by flow cytometry.

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8 protocols using anti mouse cd86 pe

Macrophage Activation and Phenotyping

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Macrophages were diluted and plated in 24-well plates and incubated for 24 h with RLPa-2, LPS, and IFN-γ as described in Section 3.7.3. Cells were collected with a cell scraper, washed with phosphate buffered saline (PBS), stained with anti-mouse CD80-FITC (1 µg/mL) and anti-mouse CD86-PE (1 µg/mL) antibodies (eBioscience Inc., San Diego, CA, USA) for 30 min at 4 °C, protected from light, and washed twice again with PBS. The expression of CD80 and CD86 on the cell surface was measured by flow cytometry analysis (BD Accuri^®C6 flow cytometer, San Jose, CA, USA).

Peng S., Gu P., Mao N., Yu L., Zhu T., He J., Yang Y., Liu Z, & Wang D. (2024). Structural Characterization and In Vitro Anti-Inflammatory Activity of Polysaccharides Isolated from the Fruits of Rosa laevigata. International Journal of Molecular Sciences, 25(4), 2133.

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Phenotyping Dendritic Cell Activation

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Single-cell suspensions prepared from spleens were labeled with an antibody cocktail containing anti-mouse-CD11c-PE-Cy7, anti-mouse-CD80-APC, anti-mouse-CD86-PE, anti-mouse-H-2K^b-FITC, and anti-mouse-I-A^b-Pacific Blue (all antibodies were purchased from eBioscience). The mean fluorescence intensities (MFI) of H-2K^b (MHC class I) and I-A^b (MHC class II) in the CD11c⁺CD80^highCD86^high population were determined using a BD LSR II flow cytometer and BD FACSDiva software.

Fallon J., Tighe R., Kradjian G., Guzman W., Bernhardt A., Neuteboom B., Lan Y., Sabzevari H., Schlom J, & Greiner J.W. (2014). The immunocytokine NHS-IL12 as a potential cancer therapeutic. Oncotarget, 5(7), 1869-1884.

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OVA-Induced Dendritic Cell Activation

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The following antigens, cytokines and antibodies were used in the study: OVA (≥98%, Sigma-Aldrich, St. Louis, USA); Granulocyte-macrophage Colony Stimulating Factor (GM-CSF), Recombinant Mouse IL-4 (r-Mu IL-4) (R&D, Minneapolis, USA). Isotype control antibodies (Abs) (IgG1, IgG2a or IgG2b), anti-mouse CD11c PerCP-Cy5.5, anti-Mouse CD80 PE-Cy7, anti-Mouse CD86 PE, anti-Mouse CD40 APC, anti-mouse MHC-II FITC, anti-Mouse CD4 FITC, anti-mouse CD62L PE, anti-mouse CD25 BV421, anti-mouse IL-4 PerCP-Cy5.5, Anti-Mouse IFN-γ PerCP-Cy5.5, Anti-Mouse Foxp3 APC, Anti-Mouse CD3e (eBioscience, San Diego, USA); Mouse Naïve CD4⁺ T cell isolated kit (Stemcell, Carlsbad, USA). Red Blood Cell Lysis Buffer (Beyotime, Shanghai, China)

Wang Y., Yu Z., Zhou Y., Zhu Y., Wang J., Fu J., Yuan Y., Chen S., Wang Y., Yu W., Gao P., Zhu W., Cheng Q., Cho S.H., Kong W, & Chen J. (2020). Different doses of ovalbumin exposure on dendritic cells determine their genetic/epigenetic regulation and T cell differentiation. Aging (Albany NY), 12(24), 25432-25451.

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Quantifying M1 and M2 Macrophages

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After different stimulations, single cell suspensions of BMDMs were prepared in flow buffer and incubated with antibodies. Anti-mouse CD86 PE (eBioscience, California, USA, Cat# 12-0862) was used to identify M1 macrophages while anti-mouse CD206 FITC (BioLegend, California, USA, Cat# 141704) was used to identify M2 macrophages. Results were acquired with a BD Canto II flow cytometer (BD Biosciences, USA) and analyzed by the FlowJo software (Tree Star, USA).

Zhou Z., Tang Y., Jin X., Chen C., Lu Y., Liu L, & Shen C. (2016). Metformin Inhibits Advanced Glycation End Products-Induced Inflammatory Response in Murine Macrophages Partly through AMPK Activation and RAGE/NFκB Pathway Suppression. Journal of Diabetes Research, 2016, 4847812.

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Phenotypic Analysis of Mouse and Human Macrophages

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After isolation, the peritoneal- and bone marrow-derived macrophages from mice were culture and analyzed by FACS using the following antibodies: F4/80-APC, CD206-Alexa Fluor 488, CD80-PE, anti-mouse CD86-PE, CD11c-APC (eBioscience). Human macrophages were analyzed following Fc receptor blocking with Human TruStain FcX™ (Biolegend), using antibodies to CD64-APC, CD163-PE-Cyanine7, CD206-Alexa Fluor 488, and CD68-PE (eBioscience). Data were collected by LSR II flow cytometry (BD) and analyzed with FlowJo software (FlowJo Enterprises, Ashland OR).

Tripathi M., Nandana S., Billet S., Cavassani K.A., Mishra R., Chung L.W., Posadas E.M, & Bhowmick N.A. (2017). Modulation of cabozantinib efficacy by the prostate tumor microenvironment. Oncotarget, 8(50), 87891-87902.

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Phenotypic Characterization of Mouse iPS-MSCs

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The samples of mouse iPS-MSCs (passage 5-7) and mouse BMSCs (Cyagen) were analyzed by flow cytometry with a BD FACS Canto II flow cytometer. Compensation was set using BD Comp Beads. Cells were tested for MSC positive markers, CD29 PE (eBioscience, San Diego, CA), anti-mouse CD44 FITC (eBioscience), anti-mouse CD73 PE (R&D Systems, Minneapolis, MN), anti-mouse CD90 PE (R&D) and MSC negative markers, anti-mouse CD31 FITC (eBioscience), anti-mouse CD45 FITC (eBioscience), anti-mouse CD86 PE (eBioscience). Data were analyzed using FACS Diva software (BD Biosciences,
Franklin Lakes, New Jersey) and Flowjo™ data analysis package (Tree Star, Inc., Ashland, OR).

Hontani K., Onodera T., Terashima M., Momma D., Matsuoka M., Baba R., Joutoku Z., Matsubara S., Homan K., Hishimura R., Xu L, & Iwasaki N. (2019). Chondrogenic differentiation of mouse induced pluripotent stem cells using the three-dimensional culture with ultra-purified alginate gel. Journal of biomedical materials research. Part A, 107(5).

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Immunophenotyping of Tumor-Challenged Mice

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Tumor-challenged mice were treated as described earlier, and on day 10 after the first treatment, mice from each group (n=3) were euthanized and the cells from the spleens, tumor-draining lymph nodes (TDLNs), and tumors were surgically removed and used for MDSC, Treg, DC, and CD4⁺ and CD8⁺T-cell quantification by flow cytometry. A single cell suspension of spleens and TDLNs was prepared by filtration through a 300-gauge mesh. Tumor suspensions were prepared as described previously, with modifications.28 (link) The cell suspensions were then stained at 4 degree for 30 min using the following antibodies: FITC anti-mouse CD11b, PE anti-mouse Gr-1, FITC anti-mouse CD4, PE anti-mouse FOXP3, FITC anti-mouse CD11c, PE anti-mouse CD86, PE-cy5 anti-mouse CD3, FITC anti-mouse CD4, PE anti-mouse CD8, and the corresponding isotype control antibodies (all monoclonal antibodies were obtained from eBioscience). After washing with PBS, the cells were fixed with 10% formaldehyde. The cell frequency was determined using BD FACSCalibur. The samples were analyzed using FlowJo software (FlowJo, Ashland, OR, USA).

Yin L., Zhao C., Han J., Li Z., Zhen Y., Xiao R., Xu Z, & Sun Y. (2017). Antitumor effects of oncolytic herpes simplex virus type 2 against colorectal cancer in vitro and in vivo. Therapeutics and Clinical Risk Management, 13, 117-130.

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Trichinella spiralis Infection Murine Model

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Trichinella spiralis (ISS534) were obtained from specific pathogen-free female inbred Wistar rats (220.0 ± 20.0 g), maintained by serial oral passage. Eight-week-old BALB/c and C57BL/6 male mice (22.0 ± 2.0 g) were housed under proper care and specific pathogen-free conditions by the institutional guidelines. Wistar rats, BALB/c and C57BL/6 mice were purchased from Jilin University Experimental Animal Center (Jilin, China), with certificate No. 2016-0001 in conformity with SCXK (Jilin). The H22 hepatoma cell line was purchased from Bio-Rad Life Sciences Development Co., Ltd. (Beijing, China). FITC anti-mouse CD11c, PE Anti-Mouse MHC Class II (I-A), and PE anti-mouse CD86 were purchased from eBioscience (USA).

Ding J., Liu X., Tang B., Bai X., Wang Y., Li S., Li J., Liu M, & Wang X. (2020). Murine hepatoma treatment with mature dendritic cells stimulated by Trichinella spiralis excretory/secretory products. Parasite, 27, 47.

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