Benzonase nuclease can effectively reduce the viscosity of protein samples, remove the contamination of nucleic acid in protein samples, and has no residual protease activity. The nuclease activity reaches 1 * 106 U/mg protein. Benzonase nuclease also has many other applications, such as reducing sample processing time, increasing protein yield, more complete precipitation and supernatant separation during centrifugation, more convenient centrifugation of solution (especially ultrafiltration), and improving the efficiency of chromatographic purification.
Our Benzonase nuclease includes four types: Benzonase nuclease without any label, endotoxin-free Benzonase nuclease, Strep tagII Benzonase nuclease, and endotoxin-free Strep tagII Benzonase nuclease, which can meet the needs of different downstream applications. Among them, Strep tagII Benzonase nuclease can be efficiently removed by Strep Tactin XT Resin after digestion of nucleic acid. And endotoxin-free Benzonase nucleases, with endotoxin content < 40 EU/mL (calculated by unit conversion amount as endotoxin <0.15 EU per 1000U), can be applied to the development and production of medicinal grade. In addition, the nuclease residue can be evaluated by Benzonase residue detection kit based on fluorescent probe, and the detection can be completed in 15 minutes, with a minimum of 2 ng/ml of nuclease residue detectable.
The amount of enzyme that reduced the value of △A260 by 1.0 (equivalent to the complete digestion of 37 μg DNA) within 30 minutes under the reaction conditions of 37℃ and pH 8.0 is defined as an active unit.
Note: Crude products containing a large amount of protein, cell wall and other salts can partly inhibit the activity of the enzyme, and the amount of the enzyme needs to be increased when used.
- Removal of nucleic acid contamination during protein extraction: Benzonase nuclease can effectively reduce sample viscosity and facilitate downstream operation when purifying recombinant protein or extracting protein from tissue cell samples
- Use with cell or bacterial lysate to remove nucleic acid from crude extract, reduce solution viscosity and increase protein yield
- Reduction of clotting of stored peripheral blood monocytes (PBMCs)
- Degradation of nucleic acids to facilitate the preparation of high-quality inclusion bodies before renaturation of insoluble proteins
- Effective removal of negatively charged nucleic acids on bidirectional SDS-PAGE protein samples improves protein separation and enhances 2-DE resolution
- Removal of DNA Contamination in Vaccine and Virus Sample Preparation
Benzonase nuclease has high stability and digestive nucleic acid activity under very broad conditions, which makes them compatible with a variety of cell lysates and protein extraction reagents containing a variety of ionic and nonionic detergents, reductants, etc. For example, 1 mM EDTA partially inhibits Benzonase activity, 5 mM EDTA loses 90% of activity, and 1 mM PMSF does not inhibit nuclease activity. < 0.4% Triton® X-100 has almost no effect on nuclease activity. <0.4% sodium deoxycholate can maintain 70% nuclease activity, but after exceeding this threshold, nuclease activity will decrease sharply, and 1% concentration will reduce 70%. 0.05% SDS has almost no effect on nuclease activity, while SDS concentration between 0.1% and 1% will decrease nuclease activity sharply after denaturation. Increasing the amount of nuclease can compensate for activity. In addition, the nuclease can tolerate 6M urea. When the urea concentration reaches 7M, the activity of nuclease decreases sharply after denaturation, which can also be compensated by increasing the amount of nuclease.
Removal of Benzonase
For Strep tag II Benzonase, the removal rate is >95% by Strep-Tactin XT Resin. For untagged Benzonase nuclease, the nuclease can be removed by chromatographic purification, and then Benzonase Detection Kit can be used to detect remaining nuclease. Common chromatographic purification methods include cation exchange media and anion exchange media
Stored at -20℃ for 2 years.