Cells from spleen and peripheral lymph nodes (PLN) were obtained by gentle pressure-dissociation of spleen and PLN using FACS buffer, and passed through a 40-μm nylon cell strainer, and the cells were further centrifuged at 100 g for 5 min to pellet the cells. Red blood cells were lysed, and removed by addition of 5 mL ACK lysis buffer (Lonza, Walkersville, MD), mixed briefly to re-suspend cells and incubated for 2-3 minutes at room temperature, followed by FACS buffer wash and spin steps. Cells were re-suspended in FACS buffer and incubated with Fc block at room temperature for 15 minutes. Cells were divided into individual tubes, and incubated with 100 μL of FACS buffer containing antibody cocktails against CD45-APC (clone 30-F11), mouse Ly-6G (Gr-1) Alexa Fluor 488 (clone RB6-8C5), and/or mouse CD11b-PE (clone M1/70) (all from BD Biosciences) in FACS buffer for 30 min. Samples were washed and the data was acquired by FACS Caliber flow cytometry (BD Biosciences). The data was further analyzed using FlowJo software (FlowJo LLC, Oregon). The dead cells were excluded by propidium iodide staining.
Mouse cd11b pe clone m1 70
Manufactured by BD
Mouse CD11b-PE (clone M1/70) is a fluorescently-labeled monoclonal antibody that binds to the CD11b antigen, also known as the integrin alpha M chain or Mac-1 alpha chain, expressed on the surface of various myeloid cells, including monocytes, macrophages, and granulocytes.
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2 protocols using mouse cd11b pe clone m1 70
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Isolation and Characterization of Immune Cells
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Isolation and Characterization of Immune Cells
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Cells from spleen and peripheral lymph nodes (PLN) were obtained by gentle pressure-dissociation of spleen and PLN using FACS buffer, and passed through a 40-μm nylon cell strainer, and the cells were further centrifuged at 100 g for 5 min to pellet the cells. Red blood cells were lysed, and removed by addition of 5 mL ACK lysis buffer (Lonza, Walkersville, MD), mixed briefly to re-suspend cells and incubated for 2-3 minutes at room temperature, followed by FACS buffer wash and spin steps. Cells were re-suspended in FACS buffer and incubated with Fc block at room temperature for 15 minutes. Cells were divided into individual tubes, and incubated with 100 μL of FACS buffer containing antibody cocktails against CD45-APC (clone 30-F11), mouse Ly-6G (Gr-1) Alexa Fluor 488 (clone RB6-8C5), and/or mouse CD11b-PE (clone M1/70) (all from BD Biosciences) in FACS buffer for 30 min. Samples were washed and the data was acquired by FACS Caliber flow cytometry (BD Biosciences). The data was further analyzed using FlowJo software (FlowJo LLC, Oregon). The dead cells were excluded by propidium iodide staining.
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