The high local trypsin concentration in the column plus the large internal surface area across a short distance ensures almost instant binding and cleavage of proteins and antibodies at room temperature. Complete antibody digestion takes 3–6 minutes as compared to traditional methods that may require overnight digestion.
Figure 1. Capturem Trypsin digestion workflow. After initial activation of the columns with the included Capturem Trypsin Activation Buffer, the sample solution (50–800 µl) containing the native or denatured sample is passed through the spin column via two-minute centrifugation. Digestion by trypsin occurs on the column, and subsequent peptide fragments are eluted in a second two-minute centrifugation. *Denaturation/reduction/alkylation time is not included.