ELISA versus LUMINEX assay for measuring mouse metabolic hormones and cytokines: sharing the lessons I have learned

Enzyme-linked immunosorbent assay (ELISA) has long been the standard for quantitative analysis of metabolic hormones and cytokines. LUMINEX multiplex bead array assays were developed as cost- and time-effective alternatives to ELISA, but they are the only cost- and time-effective if they provide informative data. Here, I show that using half-volume of reagents for an adiponectin single-plex LUMINEX assay and a 6-plex LUMINEX xMAP mouse metabolic bead assay, produces reliable data and increases assay cost-effectiveness. I provide direct comparisons between LUMINEX assay and ELISA for quantitation of mouse leptin and insulin, and evaluate glucagon, GLP-1, IL-6, and TNFα data obtained using the 6-plex LUMINEX assay for a high-fat diet-induced obesity study. Good correlations between assays were obtained for fasting leptin and non-fasting insulin. However, the LUMINEX assay proved unsuitable for quantitating fasting insulin. ELISA proved suitable for quantitating fasting male, but not female, insulin. The LUMINEX assay gave lower values for leptin and higher values for insulin, compared with ELISA. The mouse metabolic LUMINEX assay proved unsuitable for quantitating glucagon, GLP-1, IL-6, and TNFα, due to undetectable levels in most fasting and non-fasting plasma. Overall, quantitative leptin levels were the only reliable data obtained from the mouse metabolic LUMINEX assay.