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85601
CD11b/ITGAM (M1/70) Rat mAb (PerCP-Cy5.5® Conjugate)
Antibody Conjugates

Monoclonal Antibody - CD11b/ITGAM (M1/70) Rat mAb (PerCP-Cy5.5® Conjugate), UniProt ID P11215, Entrez ID 3684 #85601

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Flow cytometric analysis of live mouse bone marrow cells using CD11b/ITGAM (M1/70) Rat mAb (PerCP-Cy5.5® Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PerCP-Cy5.5® Conjugate) #79201 (dashed line).

To Purchase # 85601S

Supporting Data

REACTIVITY H 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 0.25 μg 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, 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

CD11b/ITGAM (M1/70) Rat mAb (PerCP-Cy5.5® Conjugate) recognizes endogenous levels of total CD11b protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology:

Human

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

Cluster of differentiation molecule 11b (CD11b)/Integrin alpha M (ITGAM) is a transmembrane protein forming heterodimers that are composed of α and β subunits (1). CD11b is expressed by, and commonly used as a marker for, myeloid lineage cells, including neutrophils, monocytes, macrophages, and microglia (2). CD11b is phosphorylated at Ser1126 (cytoplasmic tail) in neutrophils. Research has shown that this phosphorylation event plays a role for leukocytes traveling from the blood stream to tissues (3). Furthermore, genome-wide association studies have linked CD11b to autoimmune diseases, such as systemic lupus erythematous (SLE) (4).

The M1/70 antibody is widely used as a marker for CD11b expression on neutrophils, macrophages, NK cells, and dendritic cells (DC) (5,6).

  1. Solovjov, D.A. et al. (2005) J Biol Chem 280, 1336-45.
  2. Murray, P.J. and Wynn, T.A. (2011) Nat Rev Immunol 11, 723-37.
  3. Fagerholm, S.C. et al. (2006) Blood 108, 3379-86.
  4. Rosetti, F. and Mayadas, T.N. (2016) Immunol Rev 269, 175-93.
  5. Finkelman, F.D. et al. (1996) J Immunol 157, 1406-14.
  6. Holmberg, L.A. et al. (1981) J Immunol 127, 1792-9.

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.
Cy and CyDye are registered trademarks of GE Healthcare.