Spectrophotometric assays for heparin and HS-like molecules. Heparin and HS were degraded with heparinase and heparitinase, respectively. Proteins in CM concentrated 10-fold and CMPEDF were digested first with subtilisin. Protease-treated CM and CMPEDF were reacted with heparinase and heparitinase. The appearance of degradation products (Δ4-hexuronate) was measured by absorbance at 235 nm (Extinction coefficient = 5500). Panels A and B show heparinase activity and panels C and D show heparitinase activity. Time course and concentration curves of enzymes using their respective substrates are shown in panels A and C. Panels B and D correspond to reactions with CM and CMPEDF.