rTrypsin N Protease, rTrypsin N, Protease-N, Mass Spec Grade, 20ug

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.

Physical Appearance: Supplied by lyophilized powder with 0.47mg HEPES; 0.05mg CaCl₂ per vial.

Molecular Weight: 29 kDa

Resuspension Buffer: 200 uLdistilled water, pH7.5

Storage Conditions: Store the lyophilized powder at-20℃ and reconstituted enzyme at -80℃ for up to 30 days

Shelf life: 24 months at-80℃

Stability: rTrypsin-N is maximally active at pH7.5

In-Solution Protein Digestion Protocol:

1, For maximum activity, .resuspend Mass Spec Grade rTrypsin-N in 200 uL 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 0.5ug/uL rTrypsin-N to a final enzyme: substrate ratio of 1:30. Mix well and incubated at 37℃ for 4 h.

Quality Control:

Purity: >99.5% rTrypsin-N peak area analyzed by HPLC at 280 nm

Specificity: <5% nonspecific cleavage with Human Serum Albumin(HSA) sample. Digested products were incubated at 37℃ for 4h, and was nonspecific cleavage was analyzed by LC-MS/MS

Activity: 152units/mg

Unit Definition: One unit is the amount of Mass spectrum Grade rTrypsin-N will hydrolyze per minutes at pH 10 at 25℃, OD366, Light path=1cm

MALDI-TOF Analysis: rTrysin-N is analyzed by MALDI-TOF, impurity peak is not found

LC-MS/MS Analysis: HSA samples were dissolved and denatured for at 37℃ 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