Quantitative Galectin-3 Measurement in Porcine Serum and Tissue

For circulating gal3 measurement, we used a pre-coated gal3 (LSBio, Seattle, WA kit #LSF32161) specific 96 well strip microplates ELISA kit. In our quantitative enzymes-linked assays, Gal3 ELISA was performed in triplicate well using pig serum samples collected on baseline and post-instrumentation. This colorimetric quantitative sandwich ELISA measures the optical density, which was normalized with pre-determined standard gal3 concentration according to the manufacturer’s recommendation [32 (link)]. The porcine myocardial tissue was incubated with Anti-Galectin-3 (Abcam, Cambridge, MA; ab76245) at a working dilution of 1 in 100, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) secondary antibody at 1 in 250 dilutions. Nuclear DNA was labeled in blue with DAPI (Invitrogen, P36931). Images were taken with an epifluorescence microscope (Leica DMi8 inverted) at 100× magnifications.