| Products/Services Used | Details | Operation |
|---|---|---|
| Recombinant Proteins | 10 μM adrenocorticotropic hormone from human (ACTH, 1–24, Genscript, China) or 100 µg mL−1 proteins were first denatured by heating at 95 °C for 5 min and then were incubated with 5 μg mL−1 of trypsin in a 5 mM NH4HCO3 buffer, pH 8, under 37 °C. | Get A Quote |
Enzymatic digestion for protein sequencing usually requires much time, and does not always result in high sequence coverage. Here we report the use of aqueous microdroplets to accelerate enzymatic reactions and, in particular, to improve protein sequencing. When a room temperature aqueous solution containing 10 µM myoglobin and 5 µg mL-1 trypsin is electrosonically sprayed (-3 kV) from a homemade setup to produce tiny (∼9 µm) microdroplets, we obtain 100% sequence coverage in less than 1 ms of digestion time, in sharp contrast to 60% coverage achieved by incubating the same solution at 37 °C for 14 h followed by analysis with a commercial electrospray ionization source that produces larger... More
