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以鸡胗为材料提取结蛋白,使用高离子强度的氯化钾和碘化钾的盐酸-咪唑缓冲液除去大部份肌凝蛋白、肌动蛋白和其他可溶性蛋白,再以6M尿素-磷酸盐缓冲液溶解提取,透析处理后,经DE52纤维素柱层析纯化。纯化提取物经SDS-PAGE和负染电镜技术鉴定,其分子量为51,232D,可重聚成直径为8nm的丝状结构,符合结蛋白理化特性。CBG-250法测定其蛋白浓度达1.28mg/ml。
Extract the knot protein with chicken gizzard, remove most of myosin, actin, and other soluble proteins using high ionic strength potassium chloride and potassium iodide - imidazole buffer, and then use 6M urea-phosphate buffer Dissolution extraction, dialysis treatment, purification by DE52 cellulose column chromatography. The purified extracts were identified by SDS-PAGE and negative-stain electron microscopy. Their molecular weights were 51,232D and could be re-aggregated into filamentous structures with diameter of 8nm, which accorded with the physicochemical properties of desmin. CBG-250 method to determine the protein concentration of 1.28mg / ml.