Buy 3 and get the 4th FREE!* | Learn More >>
88154
F4/80 (BM8.1) Rat mAb (PerCP-Cy5.5® Conjugate)
Antibody Conjugates

F4/80 (BM8.1) Rat mAb (PerCP-Cy5.5® Conjugate) #88154

Reviews ()
Citations (0)
Filter:
  1. F

Flow cytometric analysis of live mouse peritoneal macrophages using F4/80 (BM8.1) Rat mAb (PerCP-Cy5.5® Conjugate) (right), compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PerCP-Cy5.5® Conjugate) #79201 (left), and co-stained with CD11b-Alexa Fluor® 488.

To Purchase # 88154S
Product # Size Price
88154S
100 µg $ 329

Supporting Data

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

Product Description

This Cell Signaling Technology antibody is conjugated to PerCP-Cy5.5® and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.125μg of antibody per test.

Application Dilutions
Flow Cytometry 1:160

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

PRINT

View >Collapse >

Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

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

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

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.

B. Immunostaining

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.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised August 2019

Protocol Id: 1504

Specificity / Sensitivity

F4/80 (BM8.1) Rat mAb (PerCP-Cy5.5® 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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cy and CyDye are registered trademarks of GE Healthcare.