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Description: The CBRM1/5 monoclonal antibody reacts with an activation-specific epitope of human Mac-1. CBRM1/5 binds a subset of Mac-1 molecules on neutrophils and monocytes after stimulation with chemoattractants or phorbol esters but does not recognize Mac-1 on resting myeloid cells. Through interactions with its ligands, Mac-1 participates in adhesive cell interactions. The epitope recognized by this mAb localizes to the I domain on the alpha chain of Mac-1 very close to the ligand binding site in a region that is widely exposed. CBRM1/5 blocks Mac-1 dependent adhesion to fibrinogen and ICAM-1 and inhibits chemoattractant-stimulated adhesion of eosinophils to the Intercellular Adhesion Molecule-1 (ICAM-1). It should be noted that low level activation may occur during processing of freshly drawn blood. Therefore the CBRM1.5 antibody may exhibit some binding to Mac-1 in these unstimulated samples. However, higher levels of Mac-1 expression are observed in activated samples when compared to unstimulated cells.
Applications Reported: This CBRM1/5 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This CBRM1/5 antibody has been pre-titrated and tested by flow cytometric analysis of resting and activated normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-822-49) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD11b (integrin alpha-M, ITGAM, integrin alpha-X, ITGAX) is a 165 kDa adhesion molecule that associates non-covalently with integrin beta-2 (CD18). The CD11b/CD18 heterodimeric complex is also known as integrin alpha-M beta-2, Mac-1, and CR3 (complement receptor 3). CD11b is expressed on the surface of monocytes/macrophages, granulocytes, activated lymphocytes, a subset of NK cells, a subset of dendritic cells, and microglia in the brain. CD11b/CD18 functions as the receptor for ICAM-1 (CD54), ICAM-2 (CD102), ICAM-4 (CD242), CD14, CD50, CD23, heparin, iC3b, fibrinogen, and Factor X -these adhesions are critical for cell-cell and cell-matrix interactions. CD11b is expressed on 8% of spleen cells, 44% of bone marrow cells, and less than 1% of thymocytes, and is commonly used as a microglial marker in nervous tissue. The expression of CD11b increases during monocyte maturation and expression levels vary on tissue macrophages. Further, peritoneal macrophages are reported to express higher levels of CD11b than splenic macrophages. Diseases associated with CD11b dysfunction include systemic lupus erythematosus 6 and ITGAM-related susceptibility to systemic lupus erythematosus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: antigen CD11b (p170); CD11 antigen-like family member B; CD11b; Cell surface glycoprotein MAC-1 subunit alpha; complement component 3 receptor 3 subunit; CR-3 alpha chain; Integrin alpha-M; integrin, alpha M (complement component 3 receptor 3 subunit); Leukocyte adhesion receptor MO1; Mac-1 alpha (Mac1A); macrophage antigen alpha polypeptide; MGC117044; Neutrophil adherence receptor; neutrophil adherence receptor alpha-M subunit
Gene Aliases: CD11B; CR3A; ITGAM; MAC-1; MAC1A; MO1A; SLEB6
UniProt ID: (Human) P11215
Entrez Gene ID: (Human) 3684
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