CD11b Monoclonal Antibody (M1/70), Brilliant Violet™ 605, eBioscience™

Product Specific Information

Description
The M1/70 monoclonal antibody reacts with mouse CD11b, the 165-170 kDa integrin alphaM. CD11b non-covalently associates with CD18 to form alphaMbeta2 integrin (Mac-1) and binds to CD54 (ICAM-1), C3bi, and fibrinogen. Mac-1 is expressed by macrophages, NK cells, granulocytes, activated lymphocytes and mouse B-1 cells in the peritoneal cavity. M1/70 is also cross-reactive to human CD11b, and can be used for the detection of this antigen on human peripheral blood monocytes, granulocytes, and a subset of NK cells. Through interactions with its ligands, CD11b participates in adhesive cell interactions.

Applications Tested
This M1/70 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.25 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Light sensitivity
This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) 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.
• Our internal testing suggests that Brilliant Violet™ 605 (BV605) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 605 nm; Laser: Violet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

Target Information

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.