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563960Image1.png

Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Rat IgM, κ Isotype Control (Cat. No. 563062; Left Panel) or BD Horizon™ BV605 Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563960; 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.

BV605_hCLA_v_SSC_v1.png

Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Rat IgM, κ Isotype Control (Cat. No. 563062; Left Panel) or BD Horizon™ BV605 Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563960; 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.

563960Image1.png BV605_hCLA_v_SSC_v1.png Orange-Cap_Violet.png

Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Rat IgM, κ Isotype Control (Cat. No. 563062; Left Panel) or BD Horizon™ BV605 Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563960; 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.

Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Rat IgM, κ Isotype Control (Cat. No. 563062; Left Panel) or BD Horizon™ BV605 Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563960; 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.

Product Details
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BD Horizon™
CLA; PSGL-1; CD162; P-selectin glycoprotein ligand 1; SELPL; SELPLG
Human (QC Testing)
Rat WI, also known as Wistar (outbred) IgM, κ
Lymphocyte-depleted Stromal Preparation of Tonsil
Flow cytometry (Routinely Tested)
5 µl
V S075
AB_2738512
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.
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Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  7. CF™ is a trademark of Biotium, Inc.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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563960 Rev. 2
Antibody Details
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HECA-452

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.

This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

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).

563960 Rev. 2
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the Violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the Yellow-Green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV605
407 nm
605 nm
563960 Rev.2
Citations & References
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View product citations for antibody "563960" on CiteAb

Development References (7)

  1. 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.
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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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563960 Rev. 2

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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.