Flow cytometric analysis of live mouse peritoneal macrophages using F4/80 (BM8.1) Rat mAb (APC-Cy7® Conjugate) (right) and co-stained with CD11b-violetFluor™ 450, compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (APC-Cy7® Conjugate) #63801 (left).
|Isotype||Rat IgG2b, kappa|
This Cell Signaling Technology antibody is conjugated to APC-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, pH7.2. 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/flowdyes for a 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 August 2019
Protocol Id: 1504
F4/80 (BM8.1) Rat mAb (APC-Cy7® Conjugate) recognizes endogenous levels of total F4/80 protein. This antibody detects an epitope within the extracellular domain.Species Reactivity:
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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