Preparation and Storage
推奨アッセイ手順
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- 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.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
関連製品
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 BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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: Blocking, Stimulation). 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: Blocking, Stimulation). 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: Immunoprecipitation, Western blot). 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: Blocking, Stimulation). 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: Blocking, Stimulation). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.