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.
- Brochure
- Biochemistry Grade
Lysyl Endopeptidase® - Mass Spectrometry Grade
Lysyl Endopeptidase®
- Features
- Specification
- BSA Digestion Analysis
- Procedure
- Product List
- Related Product List
- Related Information
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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Biochemistry Grade
Mass spectrometry grade
Related Information
Category
- Life Science
- Enzyme
- Protease
- Cell Culture
- Biopharmaceuticals Manufacturing
- Quality Control Test
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