PSGL-1/CD162 Antibody (HECA-452)

Western Blot: Cutaneous Lymphocyte Antigen (CLA) Antibody (HECA-452) [NB100-78039] - Analysis of human tonsil (1), human spleen (2), and human lymph node (3) tissue using CD162 antibody at a concentration of 2 ug/ml.

Immunohistochemistry-Paraffin: Cutaneous Lymphocyte Antigen (CLA) Antibody (HECA-452) [NB100-78039] - IHC analysis of formalin-fixed paraffin-embedded tissue section of human breast adenocarcinoma using CLA antibody (clone HECA-452) at 5 ug/ml concentration. The antibody generated a membrane-cytoplasmic staining in the cancerous cells of breast adenocarcinoma. Note:- Besides lymphocytes, CLA has been shown to be present on cells of normal endometrium as well as breast cancer tissues (PMID 19625313 and 22970241).

Flow (Cell Surface): Cutaneous Lymphocyte Antigen (CLA) Antibody (HECA-452) [NB100-78039] - A cell surface stain was performed on CD3+ hPBMCs with Cutaneous Lymphocyte Antigen (CLA) antibody (HECA-452) NB100-78039 and a matched isotype control NBP2-31379. Cells were incubated in an antibody dilution of 1 ug/mL for 20 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody [F0101B, R&D Systems]. A co-stain was performed using CD3 antibody NBP2-25186APC.

Flow Cytometry: Cutaneous Lymphocyte Antigen (CLA) Antibody (HECA-452) [NB100-78039] - Human peripheral blood lymphocytes stained with purified HECA-452, followed by anti-mouse IgG FITC

Flow (Cell Surface): Cutaneous Lymphocyte Antigen (CLA) Antibody (HECA-452) [NB100-78039] - A cell surface stain was performed on CD14+ hPBMCs with Cutaneous Lymphocyte Antigen (CLA) antibody (HECA-452) NB100-78039 and a matched isotype control NBP2-31379. Cells were incubated in an antibody dilution of 1 ug/mL for 20 minutes at room temperature, followed by rat-Dylight 488 conjugated secondary antibody. A co-stain was performed using CD14 antibody NB100-77758AF488.

• Reactivity: Hu, Mu
• Applications: WB, ELISA, Flow, Flow-CS, ICC/IF, IHC, IHC-Fr, IHC-P, CyTOF-ready
• Clone: HECA-452
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

PSGL-1/CD162 Antibody (HECA-452) Summary

• Immunogen: Human tonsil stroma
• Specificity: Cutaneous Lymphocyte Antigen
• Isotype: IgM Kappa
• Clonality: Monoclonal
• Host: Rat
• Gene: SELPLG
• Purity: IgM purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/ml
  • ELISA
  • Flow Cytometry < = 2 ug/10^6 cells in 100 ul
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • CyTOF-ready
  • Flow (Cell Surface)
• Application Notes: This antibody is Cytof ready. Use in ICC/IF was reported in scientific literature (PMID: 1693467).
• Control:

  SR-BI Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: IgM purified

Notes

CLA antibody clone HECA-452 is known to recognize the cutaneous lymphocyte-associated antigen / CLA which is a specialized glycosylated form of P-selectin glycoprotein ligand-1/PSGL-1. Besides lymphocytes, this clone has been shown to react with normal endometrium as well as breast cancer tissues (PMID 19625313 and 22970241).

Alternate Names for PSGL-1/CD162 Antibody (HECA-452)

• CD162 antigen
• CD162
• P-selectin glycoprotein ligand 1
• PSGL1
• PSGL-1
• PSGL-1cutaneous lymphocyte-associated associated antigen
• selectin P ligandCLA
• SELPLG

Background

Cutaneous Lymphocyte Antigen (CLA) is a single-pass type I membrane homodimer protein (140kDa) that collects in normal and inflamed skin. It is expressed on T lymphocytes, subsets of peripheral blood memory T cells, Natural Killer cells, memory B cells and dendritic cells, as well as on monocytes and granulocytes. CLA is a carbohydrate epitope of sialic acid and fucose-modified P-selectin glycoprotein ligand-1 (PSGL-1). CLA is a subset ligand for E-selectin, P-selectin, and L-selectin. It plays a role in leukocyte tethering and rolling over vascular surfaces during inflammation. Treatment of activated HUVEC cells with HECA-452 antibody inhibits lymphocyte adhesion. The HECA-452 antibody is cross-reactive with mouse CLA and is appropriate for staining formalin-fixed paraffin embedded tissue sections.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PSGL-1/CD162 Antibody (NB100-78039)(8)

Publications using NB100-78039

• Kim CH, Cheong KA, Park CD, Lee AY. Therapeutic Effects of Combination Using Glucosamine Plus Tacrolimus (FK-506) on The Development of Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice. Scand J Immunol;75(5):471-8. 2012 May. [PMID: 22023698] (IHC, Mouse)
• Yamanaka K, Dimitroff CJ, Fuhlbrigge RC et al. Vitamins A and D are potent inhibitors of cutaneous lymphocyte-associated antigen expression. J Allergy Clin Immunol. 2008 Jan [PMID: 17910894] (WB)
• Walcheck B, Leppanen A, Cummings R et al. The monoclonal antibody CHO-131 binds to a core 2 O-glycan terminated with sialyl-Lewis x, which is a functional glycan ligand for P-selectin. Blood. 2002 Jun 1 [PMID: 12010808] (ELISA, FLOW)
• Borges E, Pendl G, Eytner R et al. The binding of T cell-expressed P-selectin glycoprotein ligand-1 to E- and P-selectin is differentially regulated. J Biol Chem. 1997 Nov. [PMID: 9353350]
• 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 Jan. [PMID: 3276207]
• 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 Dec. [PMID: 1720810]
• Picker LJ, Kishimoto TK, Smith CW et al. ELAM-1 is an adhesion molecule for skin-homing T cells. Nature. 1991 Feb. [PMID: 1705666]
• Picker LJ, Michie SA, Rott LS, Butcher EC. A unique phenotype of skin-associated lymphocytes in humans. Preferential expression of the HECA-452 epitope by benign and malignant T cells at cutaneous sites. Am J Pathol. 1990 May [PMID: 1693467] (FLOW, WB, IHC)

Bioinformatics

• Gene Symbol: SELPLG