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BenzNuclease DNA and RNA Nuclease (Ultra Pure,Protease Free)

Details for Product No. ABIN2180647, Supplier: Log in to see
Gene
Species
Serratia marcescens
Host
Escherichia coli (E. coli)
Antibody Type
Recombinant
Biological Activity
Active
Application
DNA Modification (DNA Mod)
Options
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Specificity 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 ℃, which corresponds approximately to complete digestion of 37 μg DNA. Note that 1 KU=1000 units.
Characteristics N/A
Purity >99 % as determined by SDS-PAGE reduced BenzNuclease.
Endotoxin Level N/A
Grade Ultra Pure Grade
ProductDetails: Biological Activity Comment Effect of Conditions on Activity:
BenzNuclease is functional between pH 6 and 10 (optimal at pH8 - 8.5) , and from 0℃ to 42 ℃ (optimal at 35 ℃ - 42 ℃). Mg2+ (1-2 mM) is required for enzyme activity. 1 mM EDTA reduced the activity by 30% in the presence of 1 mM MgCl2; 0.1 M EDTA eliminated all enzyme activity. In the presence of 1 mM MgCl2, enzyme levels were reduced 75% by 0.1 M CaCl2 or 1 M NaCl. Under standard assay conditions, 1 mM iodoacetate had no effect on the enzymatic rate, whereas 1 mM mercaptoethanol and maleic acid reduced the activity by only 5 to 10%. 10 mM pChloromercurybenzoate completely inactivates the enzyme, while 0.64 M beta-mercaptoethanol in the presence of 2 M urea causes only partial inactivation of the enzyme. 4 or 7 M Urea increases the enzyme activity.

Removal of BenzNuclease:
BenzNuclease contain no “Tag” and used in downstream processing can be removed by various purification methods according to the purification strategy for the target protein.
Application Notes 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 twodimensional gel electrophoresis), Viscosity reduction in protein extracts.
Reagent Preparation

Reaction buffer:50 mM Tris-HCl, 1 mM MgCl2, pH 8.0 (In the case of extensive dilution before use, carrier protein such as 0.1 mg/ml HSA or BSA is generally recommended to avoid any enzyme loss from surface adsorption)
DNA Substrate: 1 mg/ml salmon sperm DNA is dissolved overnight at 4 ℃, in reaction buffer, and is then sonicated on ice to obtain a homogenous solution.
Enzyme: Different dilution of nuclease with reaction buffer
Stop reagent: Trichloroacetic acid (TCA)

Assay Procedure

Measurement of activity: The activity of any unknown nuclease can be determined from a single measurement by means of the standard curve. The specific activity of BenzNuclease is >1.5 x 10 e6 unit/mg protein.

Calculation of Results

Standard curve establishment:
- 400 μl substrate + 100 μl enzyme of known activity = 500 μl mixture
- Incubate the mixture at 37℃ for 30 min.
- Stop the reaction by addition of 400 μl cold TCA and incubate on ice for 10 min.
- Centrifuge at 8500 g for 5 min.
- Measure the absorbance of supernatant at 260 nm.
- Lot a standard curve with nuclease of known activities for each set of measurements.

Restrictions For Research Use only
Format Lyophilized
Reconstitution Please see Certificate of Analysis for specific instructions. For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Concentration 250 U/μL
Buffer N/A
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment No activity loss was observed after storage at: In lyophilized state for 1 year (4 °C), After reconstitution under sterile conditions for 3 months (-70 °C).
Supplier Images
SDS-PAGE (SDS) image for BenzNuclease DNA and RNA Nuclease (Ultra Pure,Protease Free) (ABIN2180647) The purity of BenzNuclease was determined by SDS-PAGE reduced and staining overnight ...