BD Pharmingen™
Technical Data Sheet
PE Rat Anti-Human Cutaneous Lymphocyte Antigen
Product Information
563962
CLA; PSGL-1; CD162; P-selectin glycoprotein ligand 1; SELPL; SELPLG
100 tests
5 µl
HECA-452
Lymphocyte-depleted Stromal Preparation of Tonsil
Rat (WI) IgM, κ
QC Testing: Human
V S075
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09%
sodium azide.
Material Number:
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Description
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.
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.
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.
Application Notes
Application
Flow cytometry
563962 Rev. 1
Routinely Tested
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Suggested Companion Products
Catalog Number
554656
554657
553943
349202
555899
Name
Stain Buffer (FBS)
Stain Buffer (BSA)
PE Rat IgM, κ Isotype Control
BD FACS™ Lysing Solution
Lysing Buffer
Size
500 ml
500 ml
0.1 mg
100 ml
100 ml
Clone
(none)
(none)
R4-22
(none)
(none)
Product Notices
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6.
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/pharmingen/protocols for technical protocols.
References
Autschbach F, Qiao L, Schürmann G, Wallich R, Moubayed P, Feller A, Meuer S. 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, Boumsell L, Gilks WR, et al, ed. Leucocyte typing V : White
cell differentiation antigens. Oxford: Oxford University Press; 1995:1508-1510. (Clone-specific: Immunohistochemistry)
Berg EL, Yoshino T, Rott LS, Robinson MK, Warnock RA, Kishimoto TK, Picker LJ, Butcher EC. 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)
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)
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)
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)
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)
Schlossman SF, Boumsell L, Gilks W, et al, ed. Leukocyte Typing V: White Cell Differentiation Antigens. New York: Oxford University Press; 1995. (Biology)
563962 Rev. 1
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