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抗DNA.RNA血清。特异性测试及其在定量原位杂交中的应用。

Anti-DNA.RNA sera. Specificity tests and application in quantitative in situ hybridization.

作者信息

Raap A K, Marijnen J G, van der Ploeg M

出版信息

Histochemistry. 1984;81(6):517-20. doi: 10.1007/BF00489529.

Abstract

The specificity of anti-nucleic acid antisera can immunocytochemically be evaluated with test systems which apply various nucleic acids immobilized to inert matrices. When using polyrA.polyrU as a model compound for dsRNA, it is important to prevent the formation of the triple stranded form polyrA.(polyrU)2. Anti-dsRNA antibodies which, when tested with the correct test system, proved to be present in an earlier described anti-DNA.RNA serum, could be removed by adsorption. By cytofluorometric comparison of the immuno-fluorescence signals obtained with the anti-dsRNA containing serum and the absorbed serum, it could be shown that the anti-dsRNA antibodies do not contribute to the specific signals measured after in situ hybridization. Repetitive incubations with the anti-DNA.RNA serum led to a considerable increase in immunofluorescence signal.

摘要

抗核酸抗血清的特异性可以通过免疫细胞化学方法,利用将各种核酸固定在惰性基质上的测试系统来评估。当使用聚腺苷酸·聚尿苷酸(polyrA.polyrU)作为双链RNA(dsRNA)的模型化合物时,防止三链形式聚腺苷酸·(聚尿苷酸)2(polyrA.(polyrU)2)的形成很重要。在用正确的测试系统检测时,先前描述的抗DNA.RNA血清中存在的抗双链RNA抗体可以通过吸附去除。通过对含抗双链RNA血清和吸附后血清所获得的免疫荧光信号进行细胞荧光测定比较,可以表明抗双链RNA抗体对原位杂交后测得的特异性信号没有贡献。用抗DNA.RNA血清进行重复孵育导致免疫荧光信号显著增加。

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