||The action of endoglycosidase on the latent 58-kDa form produces 42/40-kDa species. This indicates that MMP8 is an N-linked, complex glycoprotein that appears to be glycosylated at multiple sites.
||Human neutrophil collagenase (HNC) has been purified from extracts of fresh and outdated buffy coats and from exudates of phorbol myristate acetate-stimulated neutrophils. The MMP8 present in the starting material can either be latent or active, or have an app. relative molecular mass of 75-kDa and/or 58-kDa. The rather complex pattern of activation of the latent 58-kDa and 75-kDa species by trypsin, organomercurials and oxidants has been investigated. MMP8 was shown to preferentially hydrolyze type I over type II, and type III collagens in solution and to be a glycoprotein that contains complex N-linked oligosaccharides leading to multiple forms of MMP8 in SDS-PAGE.
||Human neutrophil granulocytes (Buffy Coat)
||Liquid, in 50 mM Tris-HCl, pH 7; 200 mM NaCl; 5 mM CaCl2; 1 μM ZnCl2; 0.05% Brij 35; 0,05% NaN3.
||MMP8 is very stable if aliquoted and stored (prevents auto-activation) at -70°C. Repeated freezing and thawing should be avoided.