Insulin Secretion in INS-1E Cells

Insulin secretion was measured as described (22 (link)). Briefly, INS-1E cells were washed with modified Krebs-Ringer bicarbonate HEPES solution (KRB), incubated for 30 min with KRB without glucose, and insulin secretion was induced by 30 min incubation with KRB containing 1.67 or 16.7 mM glucose with or without 10 μM forskolin. Insulin was measured by ELISA (Mercodia, Uppsala, Sweden) in cell-free supernatants and acid-ethanol extracted cell lysates. Total protein was measured in cell lysates using the Protein Assay Dye Reagent (Biorad).

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Abdulkarim B., Nicolino M., Igoillo-Esteve M., Daures M., Romero S., Philippi A., Senée V., Lopes M., Cunha D.A., Harding H.P., Derbois C., Bendelac N., Hattersley A.T., Eizirik D.L., Ron D., Cnop M, & Julier C. (2015). A Missense Mutation in PPP1R15B Causes a Syndrome Including Diabetes, Short Stature, and Microcephaly. Diabetes, 64(11), 3951-3962.

Publication 2015

Protein Assay Dye Reagent

Acid Assay Cell Elisa Ethanol Forskolin Glucose Insulin Insulin secretion Krebs ringer hepes bicarbonate solution Protein

Corresponding Organization : Erasmus Hospital

Other organizations : National Institute for Health Research, University of Cambridge, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, University of Exeter

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Variable analysis

independent variables

Glucose concentration (1.67 mM or 16.7 mM)
Forskolin (presence or absence of 10 μM)

dependent variables

Insulin secretion

control variables

Cells (INS-1E)
Incubation time (30 minutes)
Measurement technique (ELISA)

controls

Negative control: Incubation with KRB without glucose
Positive control: Incubation with KRB containing 16.7 mM glucose

Annotations

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