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Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood lymphocytes. Whole blood was stained with either BD Horizon™ BV421 Rat IgM, κ Isotype Control (Cat. No. 562708; Left Panel) or BD Horizon™ BV421 Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563961; Right Panel). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots show the correlated expression patterns of Cutaneous Lymphocyte Antigen (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals for gated events with the light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Rat Anti-Human Cutaneous Lymphocyte Antigen
Regulatory Status Legend
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
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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- 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.
Companion Products
The HECA-452 monoclonal antibody specifically reacts with Cutaneous Lymphocyte Associated Antigen (CLA), a carbohydrate domain shared by sialyl Lewis[x] (sLe[x]) and sialyl Lewis[a] (sLe[a]) antigens. It serves as a ligand for selectins including CD62E (E-selectin; ELAM-1). CLA is expressed on high endothelium and on lymphocytes including most T lymphocytes infiltrating cutaneous sites of inflammation. Amongst peripheral blood cells, it is expressed on monocytes and granulocytes and a subset of lymphocytes. The HECA-452 antibody is also reportedly crossreactive with the mouse CLA carbohydrate epitope that is transiently expressed by PSGL-1/CD162 on activated T cells. A number of studies suggest that CLA plays an important role in supporting leucocyte adhesive interactions and migration into extravascular tissues during inflammation.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.
Development References (7)
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Autschbach F, Qiao L, Schürmann G, et al. Immunohistochemical reactivity of adhesion structure section mAb (Subpanels 2-4) in peripheral and mucosa-associated lymphoid tissue and in Crohn's disease. In: 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; 1995:1508-1510.
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Berg EL, Yoshino T, Rott LS, et al. The cutaneous lymphocyte antigen is a skin lymphocyte homing receptor for the vascular lectin endothelial cell-leukocyte adhesion molecule 1. J Exp Med. 1991; 174(6):1461-1466. (Clone-specific: Blocking, Flow cytometry). View Reference
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Borges E, Pendl G, Eytner R, Steegmaier M, Zollner O, Vestweber D. The binding of T cell-expressed P-selectin glycoprotein ligand-1 to E- and P-selectin is differentially regulated. J Biol Chem. 1997; 272(45):28786-28792. (Clone-specific: Blocking, Flow cytometry, Western blot). View Reference
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Duijvestijn AM, Horst E, Pals ST, et al. High endothelial differentiation in human lymphoid and inflammatory tissues defined by monoclonal antibody HECA-452. Am J Pathol. 1988; 130(1):147-155. (Immunogen: Immunohistochemistry). View Reference
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Picker LJ, Kishimoto TK, Smith CW, Warnock RA, Butcher EC. ELAM-1 is an adhesion molecule for skin-homing T cells. Nature. 1991; 349(6312):796-799. (Clone-specific). View Reference
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Picker LJ, Warnock RA, Burns AR, Doerschuk CM, Berg EL, Butcher EC. The neutrophil selectin LECAM-1 presents carbohydrate ligands to the vascular selectins ELAM-1 and GMP-140. Cell. 1991; 66(5):921-933. (Biology). View Reference
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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.
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.