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The antibody was purified by affinity chromatography and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™ and unconjugated antibody.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
Additional reported applications (for the relevant formats) include: blocking of lymphocyte binding to E-selectin3, and immunohistochemistry1,2 of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections. The HECA-452 antibody cross-reacts with mouse skin homing lymphocytes4.
Application References:
1. Duijvestijn AM, et al. 1988. Am. J. Pathol. 130:147. (IHC) 2. Picker LJ, et al. 1991. Nature 349:796. (IHC) 3. Berg EL, et al. 1991. J. Exp. Med. 174:1461. 4. Borges E, et al. 1997. J. Biol. Chem. 272:28786.
Human peripheral blood lymphocytes stained with HECA-452 Pacific Blue™
Cutaneous Lymphocyte-assiciated Antigen (CLA) is a 140 kD homodimer protein recognized by a unique mAb, HECA-452. It is expressed on T lymphocytes in skin, subsets of peripheral blood memory T cells, NK cells, memory B cells and dendritic cells, as well as on monocytes, granulocytes and activated endothelial cells. CLA is a carbohydrate epitope of sialic acid and fucose-modified P-selectin glycoprotein ligand-1 (PSGL-1), a surface glycoprotein expressed on majority of peripheral blood leukocytes. CLA is a subset ligand for E-selectin, P-selectin, and L-selectin. It plays a role in memory lymphocyte homing, tethering, and rolling. Treatment of activated HUVEC cells with HECA-452 antibody inhibits lymphocyte adhesion. The HECA-452 antibody is cross-reactive with mouse CLA and is suitable for staining formalin-fixed paraffin embedded tissue sections.
T lymphocytes in skin, subsets of peripheral blood T cells, NK cells, B cells and dendritic cells, monocytes, granulocytes
Function:
Memory cells tethering and rolling
Ligand Receptor:
E-selectin, P-selectin, L-selectin
Antigen References:
1. Picker LJ, et al. 1990. Am. J. Pathol. 136:1053. 2. Berg EL, et al. 1991. J. Exp. Med. 174:1461. 3. Fuhlbrigge RC, et al. 1997. Nature 389:978. 4. Tu L, et al. 1999. J. Exp. Med. 189:241. 5. Yoshino T, et al. 1999. Cell. Immunol. 197:39. 6. Chang SE, et al. 2003. Acta Derm-Venereol. 83:162. 7. Schakel K, et al. 2002. Immunity 17:289. 8. Fuhlbrigge RC, et al. 2002. J. Immunol. 168:5645.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
Human peripheral blood lymphocytes were stained with purified CLA (clone HECA-452) (filled histogram) or rat IgM isotype control (open histogram), followed by biotinylated anti-rat IgM and Sav-PE.
