This mouse IgG1 antibody was generated against human spermatozoa and recognizes a carbohydrate epitope on the spermatozoa glycoform of CD52.
Human CD52 is a glycosylphosphatidylinositol (GPI)-anchored antigen expressed on the all lymphocytes as well as within the male genital tract, where it can be produced by epithelial cells of the distal epididymis and duct deferens. CD52 can be shed into seminal plasma and then acquired by sperm cells during their passage through the genital tract, thus it is detectable on the surface of epididymal sperm and in the ejaculate but not on either spermatogenetic cells or testicular spermatozoa. Protein core of the sperm and lymphocyte CD52 is identical and it is a product of a single copy gene located on chromosome 1. However, N-linked carbohydrate side chains (attached to Asn-3) and GPI-anchor structure are different. Monoclonal antibodies directed against sperm CD52 may cause sperm immobilisation and agglutination and affect hamster oocyte penetration suggesting an association of this molecule with fertilisation.
From the laboratory of John C. Herr, PhD, University of Virginia.
Part of The Investigator's Annexe program.
|Name:||Anti-CD52, Human S19 (MHS8-A5-A1-D7)|
|Specificity:||Recognizes carbohydrate epitope (S19) on spermatoza glycoform of CD52|
|Immunogen:||Generated against human spermatazoa|
|Purification Method:||Protein G affinity purified|
|Tested Applications:||WB, IHC, IP, IF|
|Storage:||Store at -20C|
S19 immunoreactivity with an N-linked carbohydrate epitope. A) The S19 ascites (1:10,000) reacted with SAGA-1 (~1525 kDa) on an immunoblot of untreated, methanol/chloroform-extracted human sperm proteins (lane 1). N-glycanase treatment of the sperm extract abolished S19 immunoreactivity (lane 2), indicating that the S19 mAb reacts with an N-linked carbohydrate epitope. B) Amido black stained control blot of untreated (lane 1) and Nglycanase- treated (lane 2) bovine fetuin. Enzymatic cleavage of N-linked carbohydrate side chains from fetuin was evident by increased mobility in SDS-PAGE.
Adapted from: Diekman AB, Norton EJ, Klotz KL, Westbrook VA, Shibahara H, Naaby-Hansen S, Flickinger CJ, and Herr JC. N-linked glycan of a sperm CD52 glycoform associated wiht human infertility. FASEB J. 13(11):1303-1313, (1999).
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