For high-salt protein purification workflows, SERVA Electrophoresis offers Salt Active Nuclease, a non-specific endonuclease for effective DNA removal.
Salt is an important component of various purification schemes. The presence of salt can minimize protein or virus aggregation, increase target solubility and improve target yield. High salt enables contaminating DNA to dissociate from associated proteins and become available for degradation. Some commonly used nucleases, such as Benzonase®, are inhibited by high salt concentrations.
Salt Active Nuclease (#18541.01) is a nonspecific endonuclease with optimum activity at high salt concentrations. The size of the end products from ssDNA varies from ~ 5 – 13 nt, while dsDNA is digested to around ~ 5 – 7 nt. It is active in a variety of buffers and at low temperatures. The enzyme is easily inactivated by treatment with a reducing agent, and the high pI (9.6) enables easy separation from a vast majority of protein targets. Its inactivation properties could also make the enzyme convenient for decontamination of molecular biology reagents.
The ability to maintain activity in high-salt conditions addresses a common challenge in protein purification workflows. Salt may be used to support target solubility, reduce aggregation and help release contaminating DNA from associated proteins. In these conditions, DNA becomes more accessible for degradation, while many available nucleases can be inhibited by the salt concentration. Salt Active Nuclease is designed for use where effective DNA removal is needed without moving away from a high-salt purification strategy.
Inactivation is achieved by adding reducing agents like TCEP. Removal can be achieved using cationic IEX columns. Due to its high pI (9.6), Salt Active Nuclease binds tightly to cationic columns, while anionic IEX columns are not recommended for its removal.
Together, its high-salt activity, low-temperature activity, broad pH range and straightforward inactivation offer useful practical properties for purification workflows. The enzyme’s high pI and column-binding properties can also support its separation from a wide range of protein targets after DNA degradation.
For detailed information on recommended operating conditions, DNA removal guidelines, inactivation protocols and SAN removal, download the Salt Active Nuclease Technical Note.
Salt Active Nuclease (#18541.01) is available through the Bio-Connect webshop for DNA removal in high-salt protein purification workflows. If you would like to discuss its compatibility with your purification process, buffer conditions or protein target, please feel free to contact us. We are happy to help you identify whether Salt Active Nuclease is suitable for your specific application.
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