Purification Protocol of β-mannosyltransferase from pig aorta

β-Mannosyltransferase that transfers mannose to Dol-PP-GlcNAc₂ (chitobiose lipid acceptor) from GDP-Man was purified from the microsomes of pig aorta. The purification protocol consisted of the following steps: (i) preparation of microsomal fraction (ii) selective solubilization by Nonident NP-40 nonionic detergent at a protein to detergent ratio of 3:2; (iii) anion exchange chromatography on DE-52 in the presence of 0.4% NP-40, (iv)hydroxylapatite column chromatography, (v) chromatography on HQ anion exchange column, (vi) octyl Sepharose chromatography.