Venoms from natural sources have interested scientists and laypersons with diverse focuses for centuries. Be it to generate a more efficient cure for envenomation, as a source of novel molecules for biodiscovery, or to analyze biological mechanisms of toxin resistance and evolution, to mention a few. 

The use of several in vitro, ex vivo, and in vivo methodologies, such as enzyme activity measurements, immunochemical assays, diverse cell cultures, electrophysiology, biological activity/toxicity determinations, and many others, has been of utmost importance for the analysis of venom function and antivenom improvement. The techniques used to obtain venoms and toxins are highly variable, dependent on the organism they are extracted from, and can significantly affect the research outputs.

In the last decade, the popularization and improvement of proteomic and transcriptomic analysis, among other high throughput methodologies, has revolutionized venom research and produced unprecedented quality and detailed data.

While the combination of these methodologies has provided a deep understanding of several venoms and their functions and allowed for the development of better treatments for envenomation, so much more remains to be explored. The goal of this Methods Collection is to provide an overview of the variety of methods available for the study of natural venoms and to compile techniques for antivenom evaluation and development.