Proteomics Tools

Table of Mass Spec Grade Proteases

Enzyme

Class

Cleavage Sites and Digestion Conditions

Trypsin Serine Protease Carboxyl side of arginine (R) and lysine (K)
Ammonium bicarbonate buffers (pH 8.0).
1:20 to 1:100 (w/w) enzyme to protein ratio
Digestion at 37°C for 2 hours to overnight
Endoproteinase Glu-C Serine Endoproteinase Carboxyl side of glutamate (G).
Ammonium bicarbonate buffers (pH 7.8)
Carboxyl side of glutamine (G) and asparagine (N)
Phosphate buffers (pH 4.0)
1:20 to 1:100 (w/w) enzyme to protein ratio
Digestion at 37°C for 2 hours to overnight
Pepsin Digestive Protease Carboxyl side of tyrosine (Y), tryptophan (W), phenylalanine (F), and leucine (L)
Acidic conditions (pH 2-4)
1:50 (w/w) enzyme to protein ratio
Digestion at 37°C for 6 hours to overnight
Chymotrypsin Digestive Protease Carboxyl side of tyrosine (Y), phenylalanine (F), and tryptophan (W) Ammonium bicarbonate buffers (pH 8)
1:20 to 1:100 (w/w) enzyme to protein ratio
Digestion at 37°C for 2 hours to overnight
Endoproteinase Arg-C Sulfhydryl Proteinase Carboxyl side of arginine (R) residues
Ammonium bicarbionate buffers (pH 8)
1:10 to 1:20 (w/w) enzyme to protein ratio
Digestion at 37°C overnight


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