The digestion time is approximately one hour. This could be longer for very hydrophobic proteins.
The optimum RapiGest concentration is 0.25% - 0.1%.
The incubation temperature should be 37 °C for about 10 minutes. The digestion is ten completed on ice for an additional 45 minutes.
Detailed steps from the Care and Use Manual:
Add 20 μL of 0.1% RapiGest SF solution in 50 mM NH4HCO3 and incubate at 37 °C for 10 min.
Remove excess solution and completely dry the gel pieces with a Speed Vac.
Gradually (one drop at a time) add trypsin (12.5 ng per μL in 50 mM NH4HCO3) to the gel slice until the gel re-swells. Continue to incubate on ice for 45 min.
Remove excess solution, add 20 μL of 50 mM NH4HCO3, and incubate overnight at 37 °C.