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Efluor 450 anti f4 80 bm8

Manufactured by Thermo Fisher Scientific
Sourced in United States

EFluor 450-anti-F4/80 (BM8) is a fluorescently-labeled antibody used for flow cytometric analysis. It specifically binds to the F4/80 antigen, which is expressed on the surface of mouse macrophages. The fluorochrome EFluor 450 is conjugated to the antibody, allowing detection of F4/80-positive cells in the blue excitation range.

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2 protocols using efluor 450 anti f4 80 bm8

1

Isolation and Analysis of Mouse Lung Cells

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The mouse lungs were cut into small pieces and incubated in Iscove's Modified Dulbecco's Medium (Invitrogen) containing 1mg/ml collagenase D (Roche, Mannheim, Germany) and 20 U/ml DNase I (Roche) at 37°C with shaking for 1–2 hours. After treating with Ammonium-Chloride-Potassium lysing buffer and filtering by a 70 µm nylon mesh, 1×106 cells were stained with a combination of monoclonal antibodies (mAbs) of PE-Cy5-anti-CD11b (M1/70, eBioscience), eFluor 450-anti-F4/80 (BM8, eBioscience), and Alexa Fluor 700-anti-Ly-6G (RB6-8C5, eBioscience); or a combination of mAbs of eFluor 450-anti-CD3 (17A2, eBioscience), FITC-anti-NK1.1 (PK136, BioLegend, San Diego, CA, USA), PE-anti-CD4 (RM4-5, BD), and APC-anti-CD8a (53-6.7, BD); or their relevant isotype-specific antibodies as described previously [26] (link), [28] (link). Before running samples, counting beads were added into the stained cells (Molecular Probes, Carlsbad, CA, USA). All samples were acquired on LSR II and analyzed by FlowJo software (TreeStar, Ashland, OR, USA).
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2

Flow cytometric analysis of BM-derived macrophages

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BM-derived mφs (BMDMs) were trypsinized and resuspended in flow cytometry buffer (PBS with 2% FBS). Following this, 1 × 106 cells were blocked with 0.5 μg rat anti-mouse CD16/32 (2.4G2; BD Pharmingen) in a volume of 100 μl for 30 min on ice. Cells were then stained with the following Abs: 5 μg/ml eFluor450 anti-F4/80 (BM8; eBioscience), 1.25 μg/ml PE Cy7 anti-CD115 (25-1152; eBioscience), or 1.2 μg/ml eFluor450 anti-CD11b (M1/70; eBioscience). Flow cytometry was performed with a Fortessa LSR II (BD Biosciences) and analyzed using FlowJo software.
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