Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either PE Rat IgM, κ Isotype Control (Cat. No. 553943; Left Panel) or PE Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563962; Right Manel). 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.
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Multiparameter flow cytometric analysis of Cutaneous Lymphocyte Antigen expression on human peripheral blood leucocytes. Whole blood was stained with either PE Rat IgM, κ Isotype Control (Cat. No. 553943; Left Panel) or PE Rat Anti-Human Cutaneous Lymphocyte Antigen antibody (Cat. No. 563962; Right Manel). 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
BD Pharmingen™
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_2721825
Aqueous buffered solution containing BSA, protein stabilizer, 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- 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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Rat IgM, κ Isotype Control RUO
Size
0.1 mg
Cat No.
553943
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563962 Rev. 1
Antibody Details
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.
563962 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
563962 Rev.1
Citations & References
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.
563962 Rev. 1
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.
