While many proteases are used in bottom up Mass Spectrometric (MS) analysis , Trypsin  is the protease of choice for most applications.

One significant drawback to trypsin digestion involves the long sample preparation times, which typically range from 4 hours to overnight for most protocols. Achieving efficient digestion usually requires that proteins substrates must first be unfolded either with surfactants or denaturants such as urea or guanidine. These chemical additives can have negative side effects, including protein modification, inhibition of trypsin, or incompatibility with downstream LC-MS/ MS.

Accordingly, additional steps are typically required to remove these compounds prior to analysis. In an effort to shorten the time required to prepare samples for LC-MS/MS analysis, we have developed an optimized protocol in which rapid and efficient digestion is achieved at temperatures as high as 70°C.

Using this approach

• proteolytic reaction times are dramatically shortened down to 60 - 120 minutes
• no chemical denaturants have been added, off -line sample cleanup is unnecessary, leading to shorter preparation times and diminished samples losses.

The Rapid Digestion Trypsin product is highly flexible. It is able to accommodate a variety of additives including reducing and alkylating agents. No restrictions on sample volume or substrate concentrations are mandated. Furthermore, the protocol is simple to follow, and requires no laboratory equipment beyond a heat block. Digestion is achieved completely using an in-solution approach, and since the enzyme is not immobilized on beads, the protocol does not have strict requirements for rapid shaking and off -line filtering to remove beads.

In addition to all of the benefits of this level of flexibility, we have also developed a Rapid Trypsin/Lys-C product. Like the Trypsin/Lys-C mixture, that was developed to prepare maximally efficiently proteolytic digests, particularly for complex mixtures, we have produced a follow-up companion product: The Rapid Trypsin/Lys-C mixture. Like the traditional Trypsin/Lys-C mixture, the Rapid Trypsin/Lys C mixture is meant to be used for samples that require improved reproducibility across samples.

More details and performance data