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19233
F4/80 (BM8.1) Rat mAb (APC-Cy7® Conjugate)
Antibody Conjugates

F4/80 (BM8.1) Rat mAb (APC-Cy7® Conjugate) #19233

APPLICATIONS

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

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).

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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

For optimal flow cytometry results, we recommend 0.25μg of antibody per test.

Application Dilutions
Flow Cytometry 1:80
Storage:

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.

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:

Mouse

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.

This Cell Signaling Technology antibody is conjugated to APC-Cy7® and tested in-house for direct flow cytometric analysis in mouse cells.

F4/80 (EMR1) is a heavily glycosylated G-protein-coupled receptor and is a well-established marker for mouse macrophages (1-3).

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.
Entrez-Gene Id
13733
Swiss-Prot Acc.
Q61549
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Cy and CyDye are registered trademarks of GE Healthcare

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Product # Size Price
19233S
100 µg $ 349.0