Lysyl Endopeptidase®, recombinant, Biopharmaceutical Analysis Grade (rLys-C)

This product is a recombinant Lysyl Endopeptidase® expressed in E. coli. Lysyl endopeptidase® is a serine protease that cleaves peptide bonds at the carboxy-terminus of lysine residues with high specificity.
Taking advantage of its excellent specificity, Lysyl Endopeptidase® is used for peptide fragmentation and peptide mapping for analysis of the primary structure of proteins. This product is checked residual DNA and Host cell protein for biopharmaceutical analysis.

Features

  • Expressed in E. coli
  • Checked residual DNA / Host cell protein
  • Missed cleavage rate is lower than competitor‘s Lys-C

Specification

Requirement Specification
Appearance Lyophilisate
Activity ≧2.0AU/mg
DNA residual test ≦10ng/mg
HCP assay ≦1.0μg/mg
Endotoxin testing <20EU/mg
Electrophoresis test (SDS-PAGE) to pass test

BSA Digestion Analysis

We incubated BSA with each protease for 1 hour and 18 hours.
After incubations, we analized the missed cleavage rate and number of peptides.

  Our Product Competitor's Product
Specificity
(Missed cleavage rate after 1 hour's incubation)
0% 0%
Specificity
(Missed cleavage rate after 18 hours' incubation)
10% 20%
Activity
(Number of peptides after 18 hours' incubation)
41 35

The data suggested that our rLys-C have higher activity and specificity in long time reaction than the competitor’s product.

Procedure

1. Sample preparation

①Dissolve or dilute the protein sample to be digested with 25 mmol/L Tris-HCl, 1 mmol/L EDTA,pH 8.5~9.0.
②Add Disulfide threitositol (DTT) or β-mercaptoethanol to the solubilized protein at the final concentration of 5 mmol/L.
③Incubate for 30 min at room temperature.
④Add iodoacetamide to the solubilized protein at the final concentration of 10 mmol/L.
⑤Incubate in dark for 30 min at room temperature.

2. rLys-C preparation

①Dissolve lyophilized powder of rLys-C with 100 μl 12.5 mmol/L Tris-HCl, pH 8.5~9.0.

3. Sample digestion

①Add the prepared rLys-C to sample solution according to the enzyme : protein mass ratio of 1:20~1:100, and incubate at 35℃~37℃ for 2~18 hours.
②The final concentration of 0.5~1.0v/v% trifluoroacetic acid is added to stop the reaction.

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Mass spectrometry grade

For research use or further manufacturing use only. Not for use in diagnostic procedures.

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