Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Companion Products
The M18/2 antibody specifically recognizes the common β2 chain of LFA-1 (CD11a/CD18, αLβ2 integrin), Mac-1 (CD11b/CD18, αMβ2 integrin), and gp150, 95 (CD11c/CD18, αXβ2 integrin). Expression of CD18 is limited to leukocytes, where it is widely distributed in consort with the three integrin α chains (CD11a, CD11b, and CD11c). Among splenocytes, NK cells have the highest density of CD18, and T lymphocytes express a higher density than the remaining cells. The β2 integrins are important mediators of leukocyte-endothelium interactions. It has been reported that M18/2 antibody blocks in vivo metastasis of the LB lymphoma to the spleen and that it blocks in vitro formation of aggregates of LB cells and splenocytes. However, other reports indicate that mAb M18/2 has no effect on CTL-mediated killing, adherence of C3bi-sensitized erythrocytes to Mac-1, antigen-specific binding of T cells to antigen-producing cells, or rejection of cardiac allografts. Recent in vitro studies indicate that M18/2 antibody stimulates adhesion of Mac-1 to its ligands C3bi and ICAM-1, and it stimulates adhesion of LFA-1 to ICAM-1, but it has no effect upon the interactions of LFA-1 with ICAM-2 nor ICAM-3.
The antibody was conjugated to BD Horizon™ BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (8)
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Driessens MH, van Hulten P, Zuurbier A, La Riviere G, Roos E. Inhibition and stimulation of LFA-1 and Mac-1 functions by antibodies against murine CD18. Evidence that the LFA-1 binding sites for ICAM-1, -2, and -3 are distinct. J Leukoc Biol. 1996; 60(6):758-765. (Clone-specific). View Reference
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Ishida Y, Chused TM, Murakami S, Abe R. Antigen-specific cell conjugate formation and long-lasting calcium responses in recognition of Mls cellular superantigen by cloned murine T lymphocytes. Cell Immunol. 1994; 155(2):414-427. (Clone-specific). View Reference
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Isobe M, Yagita H, Okumura K, Ihara A. Specific acceptance of cardiac allograft after treatment with antibodies to ICAM-1 and LFA-1. Science. 1992; 255(5048):1125-1127. (Biology). View Reference
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Sanchez-Madrid F, Simon P, Thompson S, Springer TA. Mapping of antigenic and functional epitopes on the alpha- and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and Mac-1. J Exp Med. 1983; 158(2):586-602. (Immunogen). View Reference
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Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Biology). View Reference
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Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology). View Reference
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Zahalka MA, Naor D. Beta 2-integrin dependent aggregate formation between LB T cell lymphoma and spleen cells: assessment of correlation with spleen invasiveness. Int Immunol. 1994; 6(6):917-924. (Clone-specific). View Reference
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Zahalka MA, Okon E, Naor D. Blocking lymphoma invasiveness with a monoclonal antibody directed against the beta-chain of the leukocyte adhesion molecule (CD18). J Immunol. 1993; 150(10):4466-4477. (Clone-specific). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
