Anti total caspase 3 rabbit antibody

Mouse pancreatic islets were isolated by collagenase digestion, and INS-1 832/13 rat insulinoma cells were cultured as previously described [25 (link)]. Static GSIS, qRT-PCR, and immunoblot analysis were performed using previously published methods [26 (link)]. Islet perifusion-based secretory profiles was measured using the Biorep Perifusion System (Biorep, Miami Lakes, FL). Twenty-four hours after isolation, 50 handpicked islets were loaded into each perifusion chamber; islets were perifused with Krebs buffer containing 2.8 mmol/L glucose for 20 min, followed by 16.7 mmol/L glucose for 40 min at a rate of 120 μL/min. Secreted insulin was measured using ELISA (Mercodia); results were normalized to total DNA content. The following primary antibodies were employed at the indicated dilutions: anti-total caspase-3 rabbit antibody, which detected both cleaved and total caspase-3 (1:1000, Cell Signaling, Danvers, MA); anti-PC1/3 rabbit antibody (1:1000, Cell Signaling); anti-Cyclin D2 rabbit antibody (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA), and anti-Actin mouse antibody (1:10,000, MP Bioscience, Santa Ana, CA).