Guinea pig anti plin2

Partially dispersed human islets were fixed in 3% paraformaldehyde for 15 min, permeabilized in 0.1% Tween PBS for 15 min, blocked with 2% BSA PBS, and incubated with primary and secondary antibodies (Ab) for 1 h each in 1% BSA PBS, followed by 1 μg/mL DAPI for 2 min. For Fig. 1A–C, islets were treated as above using Bodipy 488 (Invitrogen, Carlsbad, CA) during secondary Ab incubation. For Bodipy 558/568 C₁₂ labeling, islets were incubated in CMRL 1066 plus 0.5 mmol/L OA and 0.2 μg/μL Bodipy 558/568 C₁₂ for 72 h before immunostaining. Paraffin-embedded pancreata were analyzed as described (14 (link)). Primary Abs were guinea pig anti-PLIN5 (Progen Biotechnik, Heidelberg), guinea pig anti-PLIN2 (Fitzgerald, Acton, MA) both at 1:100, and rabbit anti-insulin (Santa Cruz Biotechnology, Santa Cruz, CA) at 1:200. Secondary Abs were goat anti-guinea pig-DyLight 488 and goat anti-rabbit-DyLight 594 (Jackson Immunology, West Grove, PA) at 1:1,000. Images were captured with an Axio Observer Z.1 fluorescent microscope (Carl Zeiss) using AxioVision Release 4.7.