Flow cytometric analysis of live mouse peritoneal macrophages using F4/80 (BM8.1) Rat mAb (PE-Cy7® Conjugate) (right), compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PE-Cy7® Conjugate) #43426 (left), and co-stained with CD11b-Alexa Fluor® 488.
|Source/Isotype||Rat IgG2b kappa|
This Cell Signaling Technology antibody is conjugated to PE-Cy7® and tested in-house for direct flow cytometric analysis in mouse cells.
For optimal flow cytometry results, we recommend 0.25μg of antibody per test.
Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Count cells using a hemocytometer or alternative method.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
posted June 2017
revised June 2020
Protocol Id: 1504
F4/80 (BM8.1) Rat mAb (PE-Cy7® Conjugate) recognizes endogenous levels of total F4/80 protein. This antibody detects an epitope within the extracellular domain.
This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.
F4/80 (EMR1) is a heavily glycosylated G-protein-coupled receptor and is a well-established marker for mouse macrophages (1-3). Expression of F4/80 has also been observed in microglia and subset populations of dendritic cells (4).
BM8.1 is widely used together with antibodies to CD115 (c-Fms), CD11b, and CD11c to identify myeloid/macrophage cells by flow cytometry.
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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cy and CyDye are registered trademarks of GE Healthcare.