Available Options:
| SKU | Unit | ||
|---|---|---|---|
 | EN32-050 | 5000 U ( 20U/μl ) | Ask for an offer |
| EN32-250 | 25000 U ( 20U/μl ) | Ask for an offer |
Saltonase, HL-Nuclease (EN32)
Saltonase is a cold-active, heat-labile recombinant endonuclease produced in E.coli. Saltonase originates from psychrophilic bacteria and effectively digests all types of DNA and RNA substrates in different buffer conditions and a broad range of temperatures. It is very active in demanding conditions, including low temperatures and environment with high salt content. These features make Saltonase extremely useful for removing undesired nucleic acids contamination during purification of proteins in laboratory and manufacturing workflows.
Features and advantages
- Highly active in a broad range of temperatures (>20% at 8 – 45°C).
- Extreme nucleolytic activity at high salt concentrations (optimal concentration range for NaCl or KCl is 0 – 1.1 M), and other buffer additives, which can significantly improve efficiency and purification yield in various workflows.
- Highly active in the typical buffers and grow media.
- Requires ≥ 1 mM Mg2+ to activate and shows a broad spectrum of pH activity (optimum at pH 7.5 – 9.0).
- Irreversible thermal inactivation at low temperature (15 min at 52°C in the presence of 1 mM DTT).
Applications
- Purification of biologics from residual nucleic acids in biopharma manufacturing.
- Purification of recombinant proteins and enzymes for research and diagnostic use.
- Removal of undesired nucleic acids contamination in molecular biology reagents in demanding systems.
- Reduction of viscosity in biological samples (during production, automation).
Unit definition
One unit (U) is defined as the amount of enzyme that causes an increase in absorbance at 260 nm of 1.0 in 30 minutes at 37°C in 50 mM Tris-HCl buffer, pH 8.0 (25°C) supplemented with 5 mM MgCl2, 0.1 mg/ml BSA and 0.5 mg/ml herring sperm DNA as a substrate.
Saltonase USAGE
- The optimal, final concentration of Saltonase in a reaction depends on several factors (level of nucleic acids contamination, incubation temperature and time, salt concentration and other compounds present in the reaction mixture). Much more Saltonase is needed for total nucleic acids removal than for viscosity reduction. The amount of Saltonase and incubation conditions have to be determined experimentally (we recommend using 5 – 100 U of Saltonase per 1 ml of reaction mixture or lysate at 20– 37°C for 30 – 60 min).
- For the optimal Saltonase activity, Mg2+ ions are required.
- Inactivation of Saltonase depends on the concentration of the reducing agent, inactivation time and temperature. We recommend inactivating Saltonase by incubation at 52°C for 15 min in the presence of reducing agents such as DTT (1 – 10 mM).
- The enzyme requires at least 1 mM DTT to be completely inactivated.
Tolerance to most popular purification buffers’ additives:
| Additives: | Tolerance level range |
|---|---|
| NaCl / KCl | 0 – 1.4 M |
| Urea tested range | 0 – 6.0 M |
| Imidazole tested range | 0 – 0.4 M |
| Ammonium sulfate tested range | 0 – 0.2 M |
| Triton X-100 tested range | 0 – 15% |
| SDS | not recommended |
HL-Nuclease retains activity in a broad range of listed additives’ concentrations (excluding SDS).
Storage buffer
20 mM Tris-HCl pH 7.5; 500 mM NaCl; 5 mM MgCl2; 50% (v/v) glycerol
Storage conditions
Store at -20°C in a freezer without a defrost cycle.
For long-term storage place at -80°C freezer.
Stability
Saltonase is stable at -20°C for at least 2 years.
No loss in activity was observed after 11 days of incubation at 37°C or 30 hours at 50°C.
Saltonase does not lose its activity at least ten successive freeze/thaw cycles.
Shipping conditions
Shipping on blue/dry ice.
Quality control Saltonase
- The purity >90% determined by densitometry of SDS-PAGE.
- Undetectable proteolytic activity.