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尝试对在无细胞系统中合成的促黄体生成素释放激素前体进行免疫沉淀。

Attempts to immunoprecipitate the LHRH precursor synthesized in cell free systems.

作者信息

Cohen S, Charli J L, Díaz de León L, Millar R P, Arimura A, Morrison M R, Joseph-Bravo P

出版信息

Brain Res Bull. 1986 Mar;16(3):309-14. doi: 10.1016/0361-9230(86)90050-x.

Abstract

In order to determine the molecular weight of the Luteinizing Hormone Releasing Hormone (LHRH) precursor, poly(A)-RNA from rat hypothalami and human placenta were translated in two mRNA dependent cell free translation systems. Total translation products were immunoprecipitated with two antisera that recognized LHRH high molecular weight forms. After SDS-polyacrylamide slab gel electrophoretic analysis of the immunoprecipitated material and fluorography, we detected in both tissues a protein of 50,000 daltons with the No. 1076 antiserum. This peptide was not immunoprecipitated by the No. 743 anti-LHRH antiserum or by non-immune rabbit serum. However, this protein was not displaced by excess LHRH added during the immunoprecipitation and seemed to be present in species where LHRH has not been reported. These data demonstrated that the LHRH mRNA is present in very low amounts in hypothalamus or placenta and that the sensitivity of the assay is not high enough to recognize it.

摘要

为了确定促黄体生成激素释放激素(LHRH)前体的分子量,在两种依赖mRNA的无细胞翻译系统中翻译了来自大鼠下丘脑和人胎盘的聚腺苷酸(poly(A))-RNA。用两种识别LHRH高分子量形式的抗血清对总翻译产物进行免疫沉淀。对免疫沉淀物质进行SDS-聚丙烯酰胺平板凝胶电泳分析和荧光自显影后,我们用1076号抗血清在两种组织中检测到一种50,000道尔顿的蛋白质。该肽未被743号抗LHRH抗血清或非免疫兔血清免疫沉淀。然而,该蛋白质在免疫沉淀过程中不会被过量添加的LHRH取代,并且似乎存在于尚未报道LHRH的物种中。这些数据表明,LHRH mRNA在下丘脑或胎盘中的含量非常低,并且该检测方法的灵敏度不足以识别它。

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