Here we explore the essential elements of novel target identification and validation in the modern era of drug discovery. We discuss the utility of in vitro and in vivo technologies in the search and deconvolution of first-in-class targets, the opportunities each can provide when used effectively, and the challenges each represent. We consider the steps which bridge validation of a target and the initiation of drug discovery. We also explore the risks carried into the drug discovery process and how more advanced chemical matter can continue to build the validation package. Each section is supported by real-world examples in which such technologies have enabled novel target validation. We also provide a case study to illustrate target validation concepts throughout the life of a drug discovery project.