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UltraNuclease 


Cat. No. (references)

Specification
HBP000106
5KU/20μL
HBP00010750KU/200μL
HBP000108500KU/2mL
HBP0001095000KU/20mL

Description 

 UltraNuclease is a genetically engineered endonuclaese derived from Serratia marcescens, which is capable of degrade DNA or RNA, either double or single stranded, linear or circular under a wide range of condition, completely degrade nucleic acids into 5’-monophosphate oligonucleotides with 3-5 base length.

After genetic engineering modification, the product was fermented, expressed, and purified in Escherichia coli (E. coli), which reduces the viscosity of cell supernatant and cell lysate in scientific research, but also improve the purification efficiency and functional research of protein. It can also be used in the gene therapy, virus purification, vaccine production, protein and polysaccharide pharmaceutical industry as a host residue nucleic acid removal reagent.

Molecular Weight

30kDa

Isoelectric Point

6.85

Protein Purity

≥99%(SDS-PAGE & SEC-HPLC)

Specific Activity

≥1.1×106U/mg

Optimum Temperature

 37°C

Optimum pH

8.0

Protease Activity

 negative

Bioburden

<10CFU/100,000U

Residual Host- cell Protein

≤10ppm

Heavy Metal

≤10ppm

Bacterial Endotoxin

<0.25EU/1000U

Storage Buffer

20mM Tris-HCl, pH 8.0, 2mM MgCl2 , 20mM NaCl,50% Glycerol

 Transportation and Storage

 ≤0°C transportation;-20°C Storage,2 years validity (avoid freezing-thawing)

Unit Definition 

The amount of enzyme used to change the absorption value of △A260 by 1.0 within 30min at 37 °C, pH 8.0, equivalent to digested 37μg salmon sperm DNA by cutting into oligonucleotides, was defined as an active unit (U) .

Quality Control 

  • Residual Host-cell Protein: ELISA kit 
  • Protease Residues: 250KU/mL UltraNuclease reacted with substrate for 60min, no activity was detected.
  •  Bacterial Endotoxin
  • Bioburden
  • Heavy Metal: ICP-AES, HJ776-2015. 

Operation 

  • UltraNuclease activity was significantly inhibited when SDS concentration was over 0.1% or EDTA concentration was over 1mM.
  • Surfactant Triton X-100, Tween 20 and Tween 80 had no effect on nuclease properties when the concentration was under 1.5%.

 Operation

 Optimal Operation

Valid Operation

Temperature

37℃

0-45℃

pH

8.0-9.2

6.0-11.0

Mg2+

1-2mM

1-15mM

DTT

0-100mM

>100mM

2-Mercaptoethanol

0-100mM

>100mM

Monovalent   metal   ion(Na+,K+ etc.)

0-20mM

0-200mM

PO 3-4

0-10mM

0-10

Usage and Dosage 

  • Remove exogenous nucleic acid from vaccine products, reduce the risk of residual nucleic acid toxicity and improve product safety.
  • Reduce the viscosity of feed liquid caused by nucleic acid, shorten processing time and increase protein yield.
  • Remove the nucleic acid which wrapped particle (virus, inclusion body, etc.), which is conducive to the release and purification of the particle.
  • Nuclease treatment   can   improve   the resolution and recovery of the sample for column chromatography, electrophoresis and blotting analysis. 
  • In gene therapy, nucleic acid is removed toobtainpurifiedadeno-associatedviruses

Cautions

  • Storage of UltraNuclease at 4 ° C for twoweeksdoesnotaffectthebiologicalactivity, long-term storage at 4 ° C is notrecommended.
  • Avoidfreezing-thawingat-80°C,recommendedstorageat -20°C.
  • Avoid contamination by contact with otherenzymesusedinmoleculardiagnosis
  • duringapplication.
  • For your safety and health, please wear labcoatandgloves.

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