E8263

Benzonase^® Nuclease, ultrapure

≥250 units/μL, ≥99% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution, ultrapure grade

• Synonym(s): Endonuclease from Serratia marcescens
• CAS Number: 9025-65-4
• Enzyme Commission number: 3.1.30.2 (BRENDA, IUBMB)
• MDL number: MFCD00131010
• NACRES: NA.54

Properties

• biological source: Serratia marcescens
• Quality Level: 200
• recombinant: expressed in E. coli
• grade: ultrapure grade
• Assay: ≥99% (SDS-PAGE)
• form: buffered aqueous glycerol solution
• mol wt: 30 kDa
• concentration: ≥250 units/μL
• shipped in: wet ice
• storage temp.: −20°C

Description

Application

The enzyme has been used for an assay to detect the amount of Tf1 RNA that is protected from degradation by the nuclease, Benzonase. The Tf1 element of Schizosaccharomyces pombe is a long terminal repeat-containing retrotransposon that encodes functional protease, reverse transcriptase, and integrase proteins. It has also been used in the fractionation of nuclear pellet and nuclear extract obtained from cell lysates.

Ultrapure benzonase nuclease, or Endonuclease from Serratia marcescens, has been used in a study to assess the effect of sodium dodecylbenzenesulfonate and antifoaming agents on the endonuclease activity of Serratia marcescens. Endonuclease from Serratia marcescens has also been used in a study to investigate the structure of chromatin subunits.

Used for the removal of nucleic acid from protein samples.

Biochem/physiol Actions

Benzonase^® is a genetically engineered endonuclease from Serratia marcescens. The protein is a dimer of 30 kDa subunits with two essential disulfide bonds. This endonuclease attacks and degrades all forms of DNA and RNA (single stranded, double stranded, linear and circular) and is effective over a wide range of operating conditions. It completely digests nucleic acids to 5′- monophosphate terminated oligonucleotides 3 to 5 bases in length. This is ideal for the removal of nucleic acids from recombinant proteins. It can also be used for applications where complete digestion of nucleic acids is desirable. It also reduces viscosity in protein extracts and prevents cell clumping. Pre-treatment of a protein sample with this enzyme improves its resolution on 2D gel electrophoresis by eliminating any bound nucleic acids. The optimum pH for enzyme activity is found to be 8.0-9.2.

Digests native or heat-denatured DNA and RNA.

Unit Definition

One unit will digest sonicated salmon sperm DNA to acid-soluble oligonucleotides equivalent to a ΔA₂₆₀ of 1.0 in 30 min at pH 8.0 at 37 °C (reaction volume 2.625 ml).

Physical form

Solution in 50% glycerol containing 20 mM Tris HCl, pH 8.0, 2 mM MgCl₂, and 20 mM NaCl.

Legal Information

Benzonase^® Nuclease is supplied by Merck KGaA, Darmstadt, Germany and/or its affiliates.

Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable
• Personal Protective Equipment: dust mask type N95 (US), Eyeshields, Gloves