Ab133125

Islet tissue lysates were prepared using tissue protein extraction reagent (T-PER), which were then subjected to centrifugation at 4 °C with a rotation speed of 10,000 rpm for 10 min. The protein concentration was measured using BCA kit (Beyotime). Samples underwent denaturation at 95 °C for 10 min and store overnight at −80°C. The proteins separated on the gel were transferred onto PVDF membranes and after transfer, membrane containing proteins were blocked with 5% BSA for 1 h at room temperature. Then, the blots were washed effectively with TBST for 3 times and incubated with diluted primary antibodies at 4 °C overnight. The next day, blots were washed and treated with respective secondary antibodies for 1 h at room temperature and expression of the proteins of various experimental groups was obtained using enhanced chemiluminescence reagent (GE Healthcare) and Image J software (version 146; National Institutes of Health, Bethesda) was used to analyze fold-changes of protein levels. Anti-cleaved-caspase3 (1:1000; 9953S), anti-Bcl2 (1:1000; ab32124), anti-Bax (1:1000; ab32503), anti-SREBP-1c (1:1000; ab133125), anti-GLUT2 (1:1000; ab54460), anti-GAPDH (1:1000; ab181602) antibodies were obtained from Abcam.