Broadonase Nuclease is an endonuclease that has broad spectrum nuclease activity, which can degrade nucleic acid into 5'-monophosphate nucleotides of 3-5 bases. Because of its high efficiency in degrading DNA and RNA of any form (double-stranded, single-stranded, linear, cyclic), it is also called omnipotent nuclease. It has no protein cleavage activity.
Broadonase 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. Broadonase 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 Broadonase nuclease includes four types: Broadonase nuclease without any label, endotoxin-free Broadonase nuclease, Strep tagII Broadonase nuclease, and endotoxin-free Strep tagII Broadonase nuclease, which can meet the needs of different downstream applications. Among them, Strep tagII Broadonase nuclease can be efficiently removed by Strep Tactin XT Resin after digestion of nucleic acid. And endotoxin-free Broadonase 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 Broadonase 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: Broadonase 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
Broadonase 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 Broadonase 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 Broadonase
For Strep tag II Broadonase, the removal rate is >95% by Strep-Tactin XT Resin. For untagged Broadonase nuclease, the nuclease can be removed by chromatographic purification, and then Broadonase Detection Kit can be used to detect remaining nuclease. Common chromatographic purification methods include cation exchange media and anion exchange media.
The minimum shelf life is 2 years at -20°C.
Only for research and not intended for treatment of humans or animals
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