title: Purification and quantification of L-selectin-reactive GlyCAM-1 from mouse serum.
abstract: GlyCAM-1 is an endothelial-derived mucin found in the conditioned medium of organ-cultured lymph nodes and in serum. It is a ligand for L-selectin, but also interacts with P and E-selectin. It may play a role in the modulation of leukocyte trafficking into secondary lymphoid tissues as well as chronic inflammatory sites. Here we report the development of an ELISA that quantifies GlyCAM-1 in complex biological fluids such as serum. The ELISA response is linear over a concentration range of 13-800 ng/ml. The level of GlyCAM-1 in mouse serum was found to be 1.3-1.6 micrograms/ml. Using mouse serum as starting material, we report a rapid and efficient purification scheme for the isolation of biologically active GlyCAM-1. This material may now be used to address the biological functions of GlyCAM-1 and to further define its structural features.
authors.affiliation: Department of Anatomy, University of California, San Francisco 94143-0452, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: Singer
authors.forename: S D
authors.initials: SD
authors.lastname: Rosen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D019041:L-Selectin / Q000097:blood*; D051379:Mice; D008969:Molecular Sequence Data; D009077:Mucins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification*; D011485:Protein Binding / Q000276:immunology
chemical_list: D009077:Mucins; D019041:L-Selectin; C075185:sulfated glycoprotein p50