Anti cd11b alexa fluor 488

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

The Anti-CD11b Alexa Fluor® 488 is a fluorescently labeled antibody that binds to the CD11b surface antigen. It is designed for use in flow cytometry and other fluorescence-based applications to identify and characterize cells expressing CD11b.

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Alexa Fluor 488 (6806 citations) Alexa 488 (1863 citations) CD11b (360 citations) Anti-CD11b (124 citations) Alexa Fluors (21 citations) Alexa Fluor 488 anti (6 citations) Anti-mouse CD11b Alexa Fluor® 488 (2 citations)

4 protocols using anti cd11b alexa fluor 488

Multiparameter Flow Cytometry Analysis

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On day 28 p.i., mice were sacrificed and spleens were removed under aseptic conditions. MNCs were prepared as above described stained for 20 minutes at RT in 1% BSA‐PBS buffer with the following panel of antibodies: FITC‐CD4 (eBioscience, San Diego, CA, USA) and PE‐CD25 (eBioscience), Alexa Fluor 488‐anti‐CD11b (eBioscience) and PE‐CD16/32, PE‐CD206, PE‐CD11c, PE‐CD40, PE‐CD8a, PE‐CD14, PE‐CD200 (eBioscience). For intracellular staining, MNCs were stained for 20 minutes at RT in 0.3% saponin/1% BSA‐PBS buffer with the following panel of antibodies: FITC‐CD4 and PE‐IL‐10, PE‐transforming growth factor‐β (TGF‐β), PE‐IFN‐γ, PE‐IL‐17 (eBioscience), Alexa Fluor 488‐anti‐CD11b and PE‐IL‐12, PE‐chemokine receptor7 (CCR7) (eBioscience). At least 10 000 events were collected using flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) and data were analysed using CellQuest software (BD Biosciences).

Guo S., Liu C., Yu J., Chai Z., Wang Q., Mi X., Song G., Li Y., Yang P., Feng L., Xiao B, & Ma C. (2019). Nasal delivery of Fasudil‐modified immune cells exhibits therapeutic potential in experimental autoimmune encephalomyelitis. CNS Neuroscience & Therapeutics, 25(6), 783-795.

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Murine Gingival Cell Immunophenotyping

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Freshly isolated murine gingival cells were immunostained for flow cytometry analysis. 5 × 10⁵ cells were stained with 0.1 μg/mL Fixable Viability Stain 510 (FVS510) – BD Horizon™ for 15 min at room temperature to distinguish live and dead cells. After washing with PBS, cells were blocked for nonspecific binding with 5 μg/mL with CD16/32 mAb for 20 min on ice and stained with the following monoclonal antibodies (eBioscience) for 30 min at 4 °C: 1 μg/mL anti-CD90 (Thy-1.2) APC (Clone 53–2.1), 1 μg/mL anti- F4/80 Antigen eFluor® 450 (Clone BM8), 1 μg/mL anti-Ly-6G PE (clone 1A8-Ly6g), 2.5 μg/mL anti-CD11b Alexa Fluor® 488 (Clone M1/70). Cells were washed with PBS, fixed with 4% paraformaldehyde for 20 min at room temperature and kept at PBS until their acquisition by flow cytometry. Fluorescence was evaluated by acquiring 50,000 events/sample using FACSCanto II (BD Biosciences, San Jose, California, USA). Results were analyzed using the FACSDiva (BD Biosciences, San Jose, California, USA) and presented as percentage of positive events.

Johnson L., Almeida-da-Silva C.L., Takiya C.M., Figliuolo V., Rocha G.M., Weissmüller G., Scharfstein J., Coutinho-Silva R, & Ojcius D.M. (2018). Oral infection of mice with Fusobacterium nucleatum results in macrophage recruitment to the dental pulp and bone resorption. Biomedical Journal, 41(3), 184-193.

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Cd244 Knockout Mice for Immunological Studies

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C57BL/6 (WT) mice were purchased from The Jackson Laboratories (Bar Harbor, ME).
Cd244^-/- mice on the C57BL/6 background [12 (link)], were kindly provided by Dr. Raymond Welsh (University of Massachusetts Medical Center, Worcester, MA). Mice were maintained in the Translational Biomedical Research Center of the Medical College of Wisconsin and both genders were used for experiments in an age (6–12 week) and gender-matched manner. anti-CD45R-PE-Texas Red, anti-CD45R-PE, anti-GL7-FITC and anti-CD5-APC were purchased from BD Biosciences (San Jose, CA). CD21-eFluor 450, anti-CD23-PE-Cy7, anti-CD93-biotin, anti-CD4-APC-eFluor 780, anti-CD8-PE-Cy7, anti-TCRβ-FITC, anti-TCRβ-PE, anti-CD11b-Alexa Fluor 488, anti-CD244, anti-Foxp3-PE, anti-CD19-Alexa 700, anti-CD4-PE, anti-IgG-FITC and Streptavidin PE-Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-CD11b-Brilliant Violet 605, anti-CD11c-PE, anti-NK1.1-APC, anti-IgM-APC Cy7, anti-IgD-Pacific Blue, anti-CD38-Alexa Fluor 647, anti-CD138-APC and rat anti-mouse IgG_2a-biotin were purchased from Biolegend (San Diego, CA). Anti-IgM-FITC and the SBA Clonotyping System-B6/C57J-HRP were purchased from Southern Biotech (Birmingham, AL). 4-Hydroxy-3-nitrophenylacetyl (NP)-ficoll, NP-Chicken Gamma Globulin (CGG), NP(24)-PE and NP-BSA were purchased from Biosearch Technologies (Novato, CA).

Ray A., Yuan C.Y., Miller N.M., Mei H, & Dittel B.N. (2015). 2B4 Is Dispensable for T-Dependent B Cell Immune Responses, but Its Deficiency Leads to Enhanced T-Independent Responses Due to an Increase in Peritoneal Cavity B1b Cells. PLoS ONE, 10(8), e0137314.

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Murine Peritoneal Macrophage Infection

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Peritoneal macrophages were harvested from WT and β₃^−/− mice as described in Animal Models. 5×10⁵ murine peritoneal cells were seeded overnight on poly-L-lysine coated coverslips. After overnight incubation, cells were washed in cold RPMI+1% BSA and infected with dlCR2-pIX-dsRed (50,000vp/cell) on ice for 1hour. Unbound virus was removed by washing in cold RPMI. Cells were fixed in cold 95% ethanol and 5% acetic acid (10min, RT) or the infection was continued at 37°C for a further 1hour prior to washing and fixing. Fixed cells were blocked with 3% BSA for 30min and then incubated at RT for 60min in anti-CD11b-AlexaFluor488 (1:100 [eBioscience, Hatfield, UK]). Cells were stained with DAPI (1:10,000, 1mg/ml [Invitrogen]), washed and mounted onto glass slides.

Browne A., Tookman L.A., Ingemarsdotter C.K., Bouwman R.D., Pirlo K., Wang Y., McNeish I.A, & Lockley M. (2015). Pharmacological inhibition of β3 integrin reduces the inflammatory toxicities caused by oncolytic adenovirus without compromising anticancer activity. Cancer research, 75(14), 2811-2821.

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