Recombinant Asp-N, Mass Spec Grade

• Cat.No.: Asp-N-41
• Product Overview: rAsp-N protease is a recombinant expressed metalloprotease, derived from Pseudomonas fragi. that specifically hydrolyzes peptide bonds at the N-terminal of aspartic residues.

Specification

• Species: Pseudomonas fragi
• Description: This enzyme can be applied for protein structure sequence analysis or proteomics research.
• Form: Lyophilized powder with 78.8 μg Tris-HCl, 0.16 mg trehalose
• Bio-activity: 1800 unit/mg
• Molecular Mass: 22.4 kDa
• Purity: > 97 % peak area analyzed by HPLC at 280 nm.
• Storage: Store the lyophilized powder at -20 centigrade.
• EC No.: EC 3.4.24.33
• Shelf life: 12 months at -20 centigrade.
• Stability: Maximally active in the pH range 7.5-8.5.
• Usage Notes: In-Solution Protein Digestion Protocol: 1. recommend digest buffer: 20 mmol Tris, 50mmol ABC or HEPES buffer;， pH range between at 7-8. 2. 20 μg denatured protein sample in 40 μL digest buffer. 3. Add 10 μL rAsp-N to reach a final enzyme to substrate ratio of 1: 10/50 to digest the samples. Mix well and incubate at 37 centigrade for 2 hours.
• Specificity: Glucagon protein incubate with rAsp N at 37 centigrade for 2 hours, specific peptides analyzed by LC-MS/MS matched with 100 % theory peptides.
• Unit Definition: The amount of 1ug Asp N protease diluted 50 times and reacted with 0.072umol Cecropin A protein. assay condition: 50ul 20mmol Tris buffer, pH=8.0, 37 centigrade incubate for 1 hour and analysis by HPLC.
• LC-MS/MS Analysis: 50 μg Glucagon was dissolved in 50ul 50mmol Tris buffer at pH 8.0, incubate with 1ug rAsp N at 37 centigrade for 2 h. The digest was analyzed by LC-MS/MS. Experimental peptide results match the peptides generated in a theoretical peptides. Nonspecificity of rAsp-N: melittin are incubated at 37 centigrade for 1 h to detect nonspecific peptides.
• Resuspension Buffer: 20 μL ultra water