Preparation and characterization of an antiserum against purified proteoglycan link proteins from avian cartilage.
Link proteins have been purified from avian xyphoid process. Cartilage was extracted in 4.0 M guanidine hydrochloride and a link fraction (A1D5) was obtained by sequential cesium chloride centrifugation. Link proteins were separated from low buoyant density proteoglycans by chromatography on Sephacryl S-200 and polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The presence of contaminating proteoglycans at various purification steps was monitored in an enzyme-linked immunosorbent assay using antiserum against avian cartilage proteoglycan monomer (anti A1D1-1400 Vo). Antiserum generated against this purified link preparation (anti link[SDS]) was characterized for its ability to bind link proteins, proteoglycan monomer, and aggregate. The serum was specific only for link proteins when tested by an enzyme-linked immunosorbent assay. Some reactivity against proteoglycan monomer was observed in a Farr-type assay.
D000818:Animals; D000906:Antibodies; D002356:Cartilage / Q000276:immunology*; D002645:Chickens; D016326:Extracellular Matrix Proteins; D008970:Molecular Weight; D011506:Proteins / Q000276:immunology* / Q000302:isolation & purification; D011509:Proteoglycans / Q000276:immunology*; D000099249:Proteoglycan Link Protein
D000906:Antibodies; D016326:Extracellular Matrix Proteins; D011506:Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein