Glucose-stimulated insulin secretion assay was performed as previously described, with minor modifications (50). Briefly, INS 832/13 cells were washed and incubated with prewarmed Krebs-Ringer bicarbonate Hepes [KRBH; 0.5% bovine serum albumin, 129 mM NaCl, 5 mM NaHCO3, 4.8 mM KCl, 1.2 mM KH2PO2, 2.5 mM CaCl2, 1.2 mM MgSO4, and 10 mM Hepes (pH 7.4)] at 37°C for 2 hours. The supernatants were collected, and the cells were stimulated with KRBH containing either 2.8 or 25.5 mM glucose for 2 hours. After supernatants were collected, cells were lysed with acid ethanol, and insulin levels in supernatants and cell lysates were measured using STELLUX Chemi Human Total Proinsulin ELISA kits (ALPCO no. 80-PINHUT-CH01).

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.