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Purified anti-human/mouse Cutaneous Lymphocyte Antigen (CLA) Antibody

Pricing & Availability
Clone
HECA-452 (See other available formats)
Regulatory Status
RUO
Workshop
V S075
Other Names
Cutaneous Lymphocyte-associated Antigen (CLA)
Isotype
Rat IgM, κ
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Product Citations
publications
HECA-452_Pure_011812
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.
  • HECA-452_Pure_011812
    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.
  • HECA-452_PURE_CLA_Antibody_2_IHC-P_011118
    Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate Buffered 1X (1.0M, pH 7.4) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/mL of purified anti-human CLA (clone HECA-452) overnight at 4°C. On the next day, tissue was incubated with Alexa Fluor® 647 goat anti-rat IgM (clone MRM-47) antibody (red) and Alexa Fluor® 594 anti-human CD45RO (Clone UCHL1) antibody (green) for an hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was scanned with a 10X objective and stitched with MetaMorph® software.
  • HECA-452_PURE_CLA_Antibody_3_IHC-P_011118
    Human paraffin-embedded skin tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate Buffered 1X (10mM, pH 6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/mL of purified CLA (clone HECA-452) antibody at 4°C overnight. On the next day, tissue was incubated with Alexa Fluor® 647 anti-Rat IgM (clone MRM-47) antibody (red) and Alexa Fluor® 594 anti-human CD44 (clone IM7) antibody (green). The nuclei were conter-stained with DAPI (blue). The image was scanned with a 10X objective and stitched with MetaMorph® software.
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321302 100 µg £82
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Description

Cutaneous lymphocyte antigen (CLA) is a 140 kD homodimer protein recognized by a unique mAb, HECA-452. It is expressed on T cells 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 the majority of peripheral blood leukocytes. CLA is a ligand for E-selectin, P-selectin, and L-selectin. It plays a role in memory lymphocyte homing, tethering, and rolling.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

FC - Quality tested
IHC-P - Verified
IHC-F - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 2.0 µg per million cells in 100 µl volume. For immunohistochemistry formalin-fixed paraffin-embedded tissue sections, a concentration range of 5.0 - 10.0 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

The HECA-452 antibody cross-reacts with mouse skin homing lymphocytes4. Treatment of activated HUVEC cells with HECA-452 antibody inhibits lymphocyte adhesion. 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.

Application References

(PubMed link indicates BioLegend citation)
  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.
  5. Ren YL, et al. 2012. Am J Clin Pathol. 138:435. PubMed
Product Citations
  1. Rouers A, et al. 2021. Cell Rep Med. 2:100278. PubMed
  2. Ma T, et al. 2022. Elife. 11: . PubMed
  3. Ren Y, et al. 2012. Am J Clin Pathol. 138:435. PubMed
  4. Eccles JD, et al. 2020. Cell Rep. 30:351. PubMed
  5. Chng MHY, et al. 2020. Immunity. 51(6):1119-1135.e5.. PubMed
  6. NULL, et al. 2022. Cell. 185:916. PubMed
  7. Sander J et al. 2017. Immunity. 47(6):1051-1066 . PubMed
  8. Bang CH, et al. 2021. J Clin Med. 10: . PubMed
RRID
AB_492894 (BioLegend Cat. No. 321302)

Antigen Details

Structure
140 kD, carbohydrate epitope of PSGL-1
Distribution

T cells 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
Cell Type
B cells, Dendritic cells, Granulocytes, Monocytes, NK cells, T cells
Biology Area
Cell Adhesion, Cell Biology, Immunology
Molecular Family
Adhesion Molecules, CD Molecules
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.

Gene ID
6404 View all products for this Gene ID
UniProt
View information about CLA on UniProt.org
Go To Top Version: 1    Revision Date: 11/30/2012

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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.
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