Purification and quantification of L-selectin-reactive GlyCAM-1 from mouse serum.
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.
Singer
M S
MS
Department of Anatomy, University of California, San Francisco 94143-0452, USA.
Rosen
S D
SD
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
D009077:Mucins; D019041:L-Selectin; C075185:sulfated glycoprotein p50