We present a new protocol for intracellular delivery of our reagents (Figure 1). In protocol P010, (also in our resource download center), we outline a straightforward, mild and cost-effective method for the application of ubiquitin(-like) reagents in live cell experiments. Reported by Brik and co-workers, glass beads are shown to enable the direct cytosolic delivery of proteins in a process termed bead loading. Here, the glass beads induce physical disruption to cells to promote intracellular protein delivery. Importantly, treated cells exhibit a normal proliferation rate, emphasizing the low toxicity of this approach. If you are interested in live cell ubiquitin experiments, please let us know and we can supply the glass beads (UbiQ-367).

Figure 1. Intracellular protein delivery by glass bead loading.