Marshall E, Howells R E
Mol Biochem Parasitol. 1985 Jun;15(3):295-304. doi: 10.1016/0166-6851(85)90091-x.
The specificity of a range of 125I labelling techniques (Chloramine T, Iodogen, Bolton and Hunter reagent, lactoperoxidase and iodosulfanilic acid) to the surface of the filarial nematode Brugia pahangi was evaluated by autoradiography of sections of labelled worms and of dried SDS-polyacrylamide gels following electrophoresis of homogenised worm extracts. It was concluded that Bolton and Hunter reagent was not surface specific but labelled proteins throughout the body of the worm. At the light microscope level autoradiography of worms labelled using Chloramine T, Iodogen, lactoperoxidase and iodosulfanilic acid demonstrated that the 125I labelling was restricted to the worm surface. Electrophoresis and autoradiography showed that each method produced a different pattern of labelled polypeptide. A polypeptide of molecular weight 30 kDa was labelled using each method except Bolton and Hunter reagent, and appears to be a major surface component.
通过对标记虫体切片以及匀浆虫体提取物电泳后的干燥SDS-聚丙烯酰胺凝胶进行放射自显影,评估了一系列125I标记技术(氯胺T、碘甘醚、博尔顿和亨特试剂、乳过氧化物酶和碘代磺胺酸)对丝虫类线虫马来布鲁线虫表面的特异性。得出的结论是,博尔顿和亨特试剂并非表面特异性的,而是标记了虫体全身的蛋白质。在光学显微镜水平上,对用氯胺T、碘甘醚、乳过氧化物酶和碘代磺胺酸标记的虫体进行放射自显影显示,125I标记仅限于虫体表面。电泳和放射自显影表明,每种方法产生的标记多肽模式不同。除博尔顿和亨特试剂外,每种方法都标记了一条分子量为30 kDa的多肽,它似乎是一种主要的表面成分。