High-performance liquid chromatographic method for the determination of insulin synthesis in biological systems.
This paper reports a two-step high-performance liquid chromatographic procedure which permits the study of the incorporation of [3H]leucine into insulin in biological systems. The first step of the procedure was size exclusion chromatography, performed on a GPC-100 column, which was eluted with 0.1 M KH2PO4-methanol (9:1, v/v). By this step the bulk of both protein and radioactivity was separated from tritiated insulin. The second step, which employs reversed-phase chromatography on an octadecylsilyl column, permits the separation of insulin from other contaminants by means of a linear gradient of acetonitrile. This simple and reproducible method was employed to test insulin synthesis in cultured human retinoblastoma Y79 cells.
Calvaruso
G
G
Istituto di Chimica Biologica, Università di Palermo, Italy.
Tesoriere
G
G
Vento
R
R
Giuliano
M
M
Carabillò
M
M
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007328:Insulin / Q000032:analysis*; D011863:Radioimmunoassay; D012175:Retinoblastoma / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D014407:Tumor Cells, Cultured
D007328:Insulin