Proteomics Tools

Table of Mass Spec Grade Proteases

Enzyme

Class

Cleavage Sites and Digestion Conditions

TrypsinSerine ProteaseCarboxyl 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-CSerine EndoproteinaseCarboxyl 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
PepsinDigestive ProteaseCarboxyl 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
ChymotrypsinDigestive ProteaseCarboxyl 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-CSulfhydryl ProteinaseCarboxyl 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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