Homologous expression and purification of human HAX‐1 for structural studies

The human HAX‐1 protein is a ubiquitously expressed multifunctional protein that regulates various cellular processes through interactions with numerous cellular proteins and RNA. The purification protocol presented here is designed explicitly for HAX‐1 expressed in mammalian cells and yields high amounts of soluble HAX‐1 protein constructs N‐terminally fused to a cleavable superfolder GFP. This protocol will enable structural studies of HAX‐1, which is predicted to undergo posttranslational modifications and be partially disordered in the absence of a binding partner.