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Flow cytometric analysis of CD54 expression on human peripheral blood lymphocytes and monocytes. Whole blood was stained with either BD Horizon™ BV711 Mouse IgG1 κ Isotype Control (Cat. No. 563044; dashed line histograms) or BD Horizon™ BV711 Mouse Anti-Human CD54 antibody (Cat. No. 564078; solid line histograms). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes (Left Panel) or monocytes (Right Panel). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BV711 Mouse Anti-Human CD54

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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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 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
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- BD Horizon Brilliant Violet 711 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 www.bdbiosciences.com/us/s/resources for technical protocols.
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The HA58 monoclonal antibody specifically binds to CD54 which is also known as ICAM-1 (intracellular adhesion molecule-1). CD54 is an 85-110 kDa type I transmembrane glycoprotein that belongs to the immunoglobulin supergene family. A soluble form of CD54 can also be found in biological fluids. CD54 is expressed on endothelial cells and both resting (weak) and activated (moderate) lymphocytes and monocytes. CD54 is a ligand for LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18). The CD54 adhesion molecule plays roles in inflammatory and immune responses and neoplasia. This antibody (NA/LE format) blocks the mixed-lymphocyte reaction (MLR) and the purified format is suitable for staining acetone-fixed, frozen tissue sections.
The antibody was conjugated to BD Horizon™ BV711 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 711-nm. BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy™5.5 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

Development References (4)
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Tsujisaki M, Imai K, Hirata H, et al. Detection of circulating intercellular adhesion molecule-1 antigen in malignant diseases. Clin Exp Immunol. 1991; 85(1):3-8. (Immunogen: Blocking, ELISA, Flow cytometry, Immunoprecipitation, Radioimmunoassay, Western blot). View Reference
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