A7L6 is a rat monoclonal IgG2a antibody derived by fusion of X63 Ag8.653 mouse myeloma cells with spleen cells from a Fisher rat immunized with high molecular mass material derived from the Engelbreth-Holm-Swarm (EHS) tumor matrix containing laminin, entactin and HSPG.
A7L6 recognizes domain IV of the core protein of the large heparan sulphate proteoglycan or perlecan. The reactivity is independent of the galactosaminoglycan moieties. Therefore, the epitope is not sensitive to heparitinase treatment. APPLICATIONS A7L6 is useful for immunoprecipitation, immunoblotting, immunocytochemistry and immuno-histochemistry on frozen and paraffin-embedded tissues. Optimal antibody dilution should be determined by titration, recommended range is 1:25 - 1:200 for immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection reagent, and 1:100 - 1:1000 for immunoblotting applications.
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Tingbø, Kolset, Ofstad, Enersen, Hannesson: "Identification and distribution of heparan sulfate proteoglycans in the white muscle of Atlantic cod (Gadus morhua) and spotted wolffish (Anarhichas minor)." dans: Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology, Vol. 143, Issue 4, pp. 441-52, (2006) (PubMed).
Tapanadechopone, Hassell, Rigatti, Couchman: "Localization of glycosaminoglycan substitution sites on domain V of mouse perlecan." dans: Biochemical and biophysical research communications, Vol. 265, Issue 3, pp. 680-90, (2000) (PubMed).
Couchman, Ljubimov, Sthanam, Horchar, Hassell: "Antibody mapping and tissue localization of globular and cysteine-rich regions of perlecan domain III." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 43, Issue 9, pp. 955-63, (1995) (PubMed).
Ljubimov, Bartek, Couchman, Kapuller, Veselov, Kovarik, Perevoshchikov, Krutovskikh: "Distribution of individual components of basement membrane in human colon polyps and adenocarcinomas as revealed by monoclonal antibodies." dans: International journal of cancer. Journal international du cancer, Vol. 50, Issue 4, pp. 562-6, (1992) (PubMed).
Couchman, Ljubimov: "Mammalian tissue distribution of a large heparan sulfate proteoglycan detected by monoclonal antibodies." dans: Matrix (Stuttgart, Germany), Vol. 9, Issue 4, pp. 311-21, (1989) (PubMed).
Horiguchi, Couchman, Ljubimov, Yamasaki, Fine: "Distribution, ultrastructural localization, and ontogeny of the core protein of a heparan sulfate proteoglycan in human skin and other basement membranes." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 37, Issue 7, pp. 961-70, (1989) (PubMed).
Couchman: "Heterogeneous distribution of a basement membrane heparan sulfate proteoglycan in rat tissues." dans: The Journal of cell biology, Vol. 105, Issue 4, pp. 1901-16, (1987) (PubMed).
Proteoglycans are macromolecules consisting of a variety of core proteins with covalently attached one or several polysaccharide chains of the glycosaminoglycan type (heparan sulphate, heparin, chondroitin sulphate, dermatan sulphate or keratan sulphate). At least two forms of basement membrane heparan sulphate proteoglycan (HSPG) have been identified. One with a large core protein (> 400 kD) and one with a small core protein (30 kD). The large HSPG is probably the most abundant basement membrane proteoglycan. It is located predominantly in the lamina lucida, where it forms clustered aggregates and interacts with other basement membrane components to form the matrix. In addition, it also plays a critical role in attachment of cells to the basal membrane via integrin receptors.