Product Number: BE03

Description

Benzonase is a non-specific endonuclease derived from Serratia marcescens (also known as Spirulinum) that has been modified through protein engineering technology.It does not contain bacterial endotoxins expressed in prokaryotes.Functionally, it can cleave various forms of DNA and RNA, whether they are single stranded, double stranded, linear, circular, or super helical, and cut off phosphate diester bonds to produce 5 ‘- phosphate nucleotides or 5’ – phosphate oligonucleotides.Broad-spectrum nucleases have no specific requirements for nucleic acid base sequences and can cleave between any nucleotide in the chain, which can be used to remove nucleic acids from biological products.In the vaccine industry, protein and polysaccharide pharmaceutical industry, it is widely used to remove host nucleic acid residues, which can cause nucleic acid contamination of vaccines and protein products to reach the level of pics (pg).Broad-spectrum nucleases can reduce the viscosity of cell lysate by degrading nucleic acids, which helps promote solution filtration/ultrafiltration in the purification process of cell derived particles such as viruses, AAV carriers, and inclusion bodies. This shortens processing time, improves the separation efficiency of precipitation and supernatant during centrifugation, and improves chromatographic purification efficiency, ultimately achieving the effect of increasing yield and effectively improving the purity of the final product.Biological analysis application: It can be used for ELISA sample preparation, chromatography, active biphasic electrophoresis (protein profiling), and Western blot analysis to improve resolution and sample recovery.

Quality Index

The following commonly used reagents have inhibitory effects on the activity of broad-spectrum nucleases, and the loss of broad-spectrum nuclease activity is within 50% within the following concentration range.

Note

1. For your safety and health, please wear laboratory clothing and protective gloves when using.
2. The concentration of tramadol within 1.5% has no significant effect on broad-spectrum nuclease activity.
3. The activity of Benzonase Lyophilized Powder is influenced by factors such as ion concentration, reaction temperature, and pH, and it is recommended to explore the optimal concentration for initial use.

Description

Benzo Nuclease is a kind of no-specific endonuclease. The Benzo Nuclease genetically engineered endonuclease from Serratia marcescens.

Benzo Nuclease degrades all kinds of DNA and RNA (double stranded, single stranded, linear and circular and supercoil) but without proteolytic activity. It is effective over a wide range of conditions and possesses an exceptionally high specific activity.

Benzo Nuclease completely digests nucleic acids to 5’monophosphate terminated oligonucleotides 2 to 5 bases in length (below the hybridization limit), which is ideal for removal of nucleic acids from recombinant proteins, enabling compliance with FDA guidelines for nucleic acid contamination. The ability of Benzo Nuclease to rapidly hydrolyze nucleic acids makes the enzyme an excellent choice for viscosity reduction to reduce processing time and increase yields of protein. For example, the enzyme is compatible with BugBuster® and PopCulture® Protein Extraction Reagents and can therefore be added along with these reagents to eliminate viscosity and remove nucleic acids from E. coli extracts. The enzyme consists of two subunits of 30 kDa each. It is functional between pH 6 and 10 and from 0-42°C and requires 1-2mM Mg2+ for activation. The enzyme is also active in the presence of ionic and non-ionic detergents, reducing agents, PMSF (1 mM), EDTA (1 mM) and urea (relative activity depends on specific conditions). Activity is inhibited by>150 mM monovalent cations,>100 mM phosphate,>100 mM ammonium sulfate, or>100 mM guanidine HCl.

• Source: Serratia marcescens
• Molecular weight: 27. 9 KD (SDS)
• Purity: ≥90%, (Ultra≥99%)(SDS-PAGE)
• PI: 6. 85
• Optimum pH: 8. 0
• Optimum temperature: 37°C
• Cofactor: 1～10mM Mg2+
• Form: Liquid enzyme is the clear Solution(Contain Glycerin)
• Solid enzyme is the white lyophilized powder.
• Concentration: 1000 U/µl.
• Activity: ≥1000U/μl
• Specific activity: ≥1. 0x106U/mg protein
• Protease: Not detectable
• Microbial limit: aerobe<10CFU/100 KU Yeast and mold: <10CFU/100 KU
• Endotoxin test: (Gel Clot LAL Assay)<0. 25EU/1000U

Storage Buffer

20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl, 50%Glycerin

Dilution Buffer

20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl.

Unit definition

One unit is defined as the amount of enzyme that causes a ΔA260 of 1. 0 in 30 minutes, which corresponds to complete digestion of 37 µg DNA.

Application

• Its high intrinsic activity and broad substrate tolerance make the endonuclease an ideal tool in a variety of biotechnological and pharmaceutical applications:
• removal of nucleic acid from protein samples
• Elimination of nucleic acids from recombinant proteins
• Purification of protein fragments from inclusion bodies;
• Sample preparation in western blotting or two-dimensional gel electrophoresis
• Viscosity reduction in protein extracts.

Shipping and Handling

Liquid enzyme: Ship with Blue Ice, Storage at -20°C
Lyophilized powder: Ship at room temperature, storage blow 4°C

Guarantee

2 years

CAS: 9025-65-4

EC: 3.1.30.2

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