Protein microarrays are well-established as sensitive tools for proteomics. Particularly, the microspot immunoassay (MIA) platform enables a quantitative analysis of (phospho-) proteins in complex solutions (e.g. cell lysates or blood plasma) and with low consumption of samples and reagents. Despite numerous biological and clinical applications of MIAs there is currently no user-friendly open source data analysis software available with versatile options for data analysis and data visualization. Here, we introduce the open source software QuantProReloaded that is specifically designed for the analysis of data from MIA experiments.