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建立一种新的基于链霉素沉淀的PrPSc的Western blot检测方法,用终浓度为60mmol/L的链霉素处理蛋白酶K消化的PrPSc贮存液,通过离心沉淀PrPSc,用Western blot对PrPSc的链霉素富集效果进行检测。结果显示,链霉素能够与PrPSc结合形成高分子量复合物,但不影响糖基化形式。此外,基于链霉素沉淀的Western blot,无论是在低浓度或是大容积的条件下,均可显著地提高对PrPSc检测的敏感性。基于链霉素沉淀的Western blot试验是一种敏感、特异、快速及灵活的检测方法,有潜力用于脑组织、外周组织及体液中低水平的PrPSc的检测。
A new Western blot method based on streptomycin-precipitated PrPSc was established. Proteinase K-digested PrPSc stock solution was treated with streptomycin at a final concentration of 60 mmol / L, and PrPSc was precipitated by centrifugation. The PrPSc chain Mycophenolate enrichment test. The results show that streptomycin can combine with PrPSc to form high molecular weight complexes, but does not affect the glycosylation pattern. In addition, Western blotting based on streptomycin significantly increased the sensitivity of PrPSc assays, both at low concentrations and at high volumes. Western blot assay based on streptomycin precipitation is a sensitive, specific, rapid and flexible detection method, which has the potential for detection of low-level PrPSc in brain tissue, peripheral tissues and body fluids.