Starzec A, Counis R, Jutisz M
Laboratoire des Hormones Polypeptidiques, CNRS, Gif-sur-Yvette, France.
J Chromatogr. 1988 May 25;440:352-60. doi: 10.1016/s0021-9673(00)94538-2.
To study the biosynthesis in situ of lutropin (LH) subunits, anterior pituitary cells in culture were employed. The cells were incubated in the presence of [35S]methionine. Labelled polypeptides, immunologically related to alpha and LH beta subunits, were isolated by specific immunoprecipitation, then analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and revealed by fluorography. Electrophoretic analysis of immunoprecipitates is a crucial step in this methodology. It permits simultaneous purification, characterization and accurate quantitation of the radioactivity specifically incorporated into LH subunits. Also, using SDS-PAGE, it is possible to isolate and identify the different processed forms of LH subunits.
为了研究促黄体生成素（LH）亚基的原位生物合成，采用了培养的垂体前叶细胞。细胞在[35S]甲硫氨酸存在的情况下进行孵育。通过特异性免疫沉淀分离出与α和LHβ亚基免疫相关的标记多肽，然后通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）进行分析，并通过荧光自显影显示。免疫沉淀物的电泳分析是该方法中的关键步骤。它允许同时纯化、表征并准确定量特异性掺入LH亚基的放射性。此外，使用SDS-PAGE，可以分离和鉴定LH亚基的不同加工形式。
