Quantitative method for the determination of glycosaminoglycans from small quantities of glomerular basement membranes.

A simple, sensitive, and efficient method is described for the quantitative determination of glycosaminoglycans from bovine renal glomerular basement membranes. After release from glomerular basement membrane by protease treatment, the glycosaminoglycans were isolated by a modified DEAE-cellulose column chromatographic procedure. Quantitation of glycosaminoglycans was achieved by hexuronate measurements with a microadaptation method employing m-hydroxydiphenyl. This procedure has proven useful in analyzing small samples of basement membranes as little as 0.75 mg. The glycosaminoglycan was identified as heparan sulfate by cellulose acetate electrophoresis and nitrous acid treatment.