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41249
CD11b/ITGAM (M1/70) Rat mAb (APC Conjugate)
Antibody Conjugates

CD11b/ITGAM (M1/70) Rat mAb (APC Conjugate) #41249

APPLICATIONS

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

Flow cytometric analysis of live mouse bone marrow cells using CD11b/ITGAM (M1/70) Rat mAb (APC Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (APC Conjugate) #34828 (dashed line).

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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 0.125 μg 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.

CD11b/ITGAM (M1/70) Rat mAb (APC 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

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 and tested in-house for direct flow cytometric analysis in mouse cells.

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.
Entrez-Gene Id
3684
Swiss-Prot Acc.
P11215
For Research Use Only. Not For Use In Diagnostic Procedures.

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

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Product # Size Price
41249S
100 µg $ 119