Recombinant Trypsin-N, Mass Spec Grade

• Cat.No.: Trypsin-N-39

Specification

• Description: rTrypsin-N, Mass Spec Grade, is an innovative protease that specifically hydrolyzes peptide bonds at the amino side of lysine and arginine residues. rTrypsin-N can be applied for post-translational modification (PTM) proteins research and is beat for protein sequencing. Peptides produced more b ions than y ions in the mass spectrometer.
• Form: Supplied by lyophilized powder with 0.47 mg HEPES; 0.05 mg CaCl2 per vial.
• Bio-activity: 152 units/mg
• Molecular Mass: 29 kDa
• Purity: > 99.5 % rTrypsin-N peak area analyzed by HPLC at 280 nm.
• Storage: Store the lyophilized powder at -20 centigrade, and reconstituted enzyme at -80 centigrade for up to 30 days.
• Shelf life: 24 months at -80 centigrade
• Stability: rTrypsin-N is maximally active at pH 7.5.
• Usage Notes: In-Solution Protein Digestion Protocol: 1. For maximum activity, resuspend Mass Spec Grade rTrypsin-N in 200 μL distilled water, pH 7.5. 2. Use 20-50 mM HEPES solution, 0.1 % BT Surfactant, or Tris-HCl (pH 7.5) for the protein mixture cleavage (recommended). 3. Add 0.5μg/μL rTrypsin-N to a final enzyme : substrate ratio of 1: 30. Mix well and incubated at 37 centigrade for 4 h.
• Specificity: < 5 % nonspecific cleavage with Human Serum Albumin (HSA) sample. Digested products were incubated at 37 centigrade for 4 h, and was nonspecific cleavage was analyzed by LC-MS/MS.
• Unit Definition: One unit is the amount of Mass spectrum Grade rTrypsin-N will hydrolyze per minutes at pH 10 at 25 centigrade, OD366, Light path = 1 cm.
• LC-MS/MS Analysis: HSA samples were dissolved and denatured for at 37 centigrade for 1 h. The denatured HSA was diluted at pH 7.5 and incubated with rTrypsin-N for 4 h. The digest was analyzed by LC-MS/MS, and the peptides results matched the peptides generated in a theoretical digestion results of HSA by rTrypsin-N.
• Resuspension Buffer: 200 μL distilled water, pH 7.5
• MALDI-TOF Analysis: rTrysin-N is analyzed by MALDI-TOF, impurity peak is not found.