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52267
F4/80 (BM8.1) Rat mAb (FITC Conjugate)
Antibody Conjugates

F4/80 (BM8.1) Rat mAb (FITC Conjugate) #52267

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Flow cytometric analysis of live mouse peritoneal macrophages using F4/80 (BM8.1) Rat mAb (FITC Conjugate) (right), compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (FITC Conjugate) #56722 (left), and co-stained with CD11b-APC.

To Purchase # 52267S
Product # Size Price
52267S
100 µg $ 189

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Isotype Rat IgG2b, kappa

Product Description

This Cell Signaling Technology antibody is conjugated to FITC and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.5μg of antibody per test.

Application Dilutions
Flow Cytometry 1:100

Storage:

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.

Protocol

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Specificity / Sensitivity

F4/80 (BM8.1) Rat mAb (FITC Conjugate) recognizes endogenous levels of total F4/80 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

Source / Purification

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.

Background

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.

  1. Hirsch, S. et al. (1981) J Exp Med 154, 713-25.
  2. Austyn, J.M. and Gordon, S. (1981) Eur J Immunol 11, 805-15.
  3. McKnight, A.J. et al. (1996) J Biol Chem 271, 486-9.
  4. Greter, M. et al. (2015) Front Immunol 6, 249.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.