New, simple insulin-receptor assay with universal application to solubilized insulin receptors and receptors in broken and intact cells
| Autor: | Erik K. Frandsen, Robby A. Bacchus |
|---|---|
| Rok vydání: | 1987 |
| Předmět: |
Endocrinology
Diabetes and Metabolism medicine.medical_treatment Placenta Centrifugation Polyethylene glycol Polyethylene Glycols chemistry.chemical_compound Internal Medicine medicine Chemical Precipitation Humans Insulin Lymphocytes Receptor Ethanol biology Cell Membrane Receptor Insulin Insulin receptor Membrane chemistry Biochemistry Liver Cell culture biology.protein |
| Zdroj: | Diabetes. 36(3) |
| ISSN: | 0012-1797 |
| Popis: | A new, simple insulin-receptor-binding assay has been devised. The assay is based on the separation of free and receptor-bound 125I-labeled insulin in 80% ethanol. It was found that the insulin-receptor complex was fully stable at this ethanol concentration, regardless of the source of the receptor employed. The assay has been evaluated with solubilized insulin receptors and membrane-bound receptors from human placenta and porcine liver as well as intact cells with the IM-9 cell line. The assay is simple, rapid, and has large capacity. Comparisons of the ethanol-based assay to the conventionally employed assays with polyethylene glycol or microfuge centrifugation for the separation of free and bound 125I-insulin revealed large discrepancies between the assays. The ethanol-based assay always appeared to provide a better separation. Microfuge centrifugation of placental membranes precipitated ∼3% of the ethanol-precipitable insulin-receptor complex, while polyethylene glycol precipitation of solubilized insulin receptors varied between 40 and 80% of the ethanol precipitability, depending on the receptor concentration employed. |
| Databáze: | OpenAIRE |
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