Trypsin Gold, Mass Spectrometry Grade, is manufactured to provide maximum specificity. Lysine residues in the porcine trypsin are modified by reductive methylation, yielding a highly active and stable molecule that is extremely resistant to autolytic digestion. The specificity of the purified trypsin is further improved by TPCK treatment, which inactivates chymotrypsin. The treated trypsin is then purified by affinity chromatography and lyophilized to yield Trypsin Gold, Mass Spectrometry Grade.
Trypsin is often used for in-gel digestion. In this complex protein, mixtures such as cell extracts, are resolved by gel electrophoresis and the band or spot of interest is excised from the gel and digested with trypsin. The digestion products are purified and concentrated, then analyzed by mass spectrometry to determine their molecular weights. Database searches can then be performed, using the mass of the peptides to identify the protein(s) resolved on the gel.
| Component Listing for V5280 | |
| 1 x 100µg | Trypsin Gold, Mass Spectrometry Grade |
Trypsin Gold, Mass Spectrometry Grade
Other online protocols for in-gel digestions using trypsin:
» Mitchison Lab at Harvard University
» University of Minnesota MSCLS
Figure 1. Spectrogram of bovine carbonic anhydrase II digested by Trypsin Gold, Mass Spectrometry Grade. A 500ng of carbonic anhydrase II was separated by gel electrophoresis and digested with 500ng Trypsin Gold, Mass Spectrometry Grade, overnight at 37°C. The peptides generated were purified as described in sections III and V of the protocol using a PerSeptive BioSystems Voyager-DE™ MALDI-TOF system. The peak at 842.51 is due to autolysis of Trypsin.
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