A sensitive assay for active link protein from cartilage

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Abstract
A new assay for the activity of cartilage link protein is described. The method is based on the finding [Plaas, Sandy and Muir (1983) Biochem. J. 214, 855-864] that addition of link protein to [35S]sulphate-labelled proteoglycan aggregates from rabbit chondrocyte cultures resulted in the formation of link-stabilized aggregates. The percentage aggregate was found to be related linearly to the amount of purified bovine link protein added in the 20-120 ng range. The assay was used to monitor loss of link-protein activity during heat denaturation and to measure binding of link protein by purified proteoglycan monomer.