Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody.

Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody. Research Abstract Details 

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Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody. Abstract Text:

Jin Xu,Jun Cai,Simon F Peek,M Suresh,Benjamin J Darien,Jin Xu,Jun Cai,Simon F Peek,M Suresh,Benjamin J Darien,Jin Xu,Jun Cai,Simon F Peek,M Suresh,Benjamin J Darien,

P-selectin glycoprotein ligand (PSGL-1) is a widely distributed adhesion molecule that plays a critical role in regulating lymphocyte homing and leukocyte trafficking during inflammation. The lack of specific reagents for equine PSGL-1 (ePSGL-1) has prevented mechanistic studies regarding its function and regulation in the horse. We synthesized a ePSGL-1 peptide to generate a monoclonal antibody (mAb), ePL1. Using flow cytometry and Western blot, we showed that ePL1 binds specifically to ePSGL-1 in transfected mammalian cells. We also demonstrated that ePL1 binds to equine leukocytes and recognized a protein with molecular weight 165 and 280kDa under reducing and non-reducing condition, respectively, likely corresponding to ePSGL-1.Seventy percent of equine monocytes bound by both ePL1 and HECA-452, an antibody defining sLex-like carbohydrate epitope. Both ePL1 and HECA-452 recognized ePSGL-1 protein precipitated by equine P-selectin-IgG chimera. Neuraminidase treatment increased ePL1 binding and the molecular weight of ePSGL-1, O-sialoglycoprotein endopeptidase digestion and tyrosine mutation abolished ePL1 staining and recognition. The ePL1 specific binding epitope appears to be the polypeptide backbone of ePSGL-1 in the presence of tyrosine but the process is independent of sialylation modification. In conclusion, we provide evidence that this antibody can be used for cell surface staining and immune-blot analyses.

Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody. Publishing Authors By Initials

J Xu,J Cai,SF Peek,M Suresh,BJ Darien,J Xu,J Cai,SF Peek,M Suresh,BJ Darien,J Xu,J Cai,SF Peek,M Suresh,BJ Darien,

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Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Veterinary immunology and immunopathology

VOLUME: 121

Page Numbers: 144-9

Journal Abbreviation: Vet. Immunol. Immunopathol.

ISSN: 0165-2427

DAY: 16

MONTH: 09

YEAR: 2007

Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody. Information

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LANGUAGE: eng

NlmUniqueID: 8002006

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Grant and Affiliation Information for Characterization of equine P-selectin glycoprotein ligand-1 by using a specific monoclonal antibody.

AFFILIATION: Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706-1102, USA.

Country: Netherlands

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MEDLINETA: Vet Immunol Immunopathol

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