DNase I (Deoxyribonuelease I)

Cat.No.:IC-1200

Description 

DNase I is found in most cells and tissues. In mammals the pancreas is one of the best sources for the enzyme. Pancreatic DNase I was the first DNase isolated. DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5¡¯-phosphates. A tetranucleotide is the smallest average digestion product. In the presence of Mg²⁺ ions, DNase I attacks each strand of DNA independently and the cleavage sites are random. If Mn²⁺ ions are present, both DNA strands are cleaved at approximately the samesite. DNase I hydrolyzes single and double-stranded DNA and chromatin (reaction rate is restricted by DNA association with histones). DNase I is used to remove DNA from protein and nucleic acid samples, and to nick DNA as a first step to incorporate labeled bases into DNA.

Size

25mg,100mg,500mg

Activity

1500U/mg

Usage

DNase I can be used in several applications:

1.Prevents unwanted cell clumping during tissue disaggregation procedures and during cell culture since it isincluded in the medium.Has been proven to be non-cytotoxic in concentrations up to 1 mg/ml.
2.Tissue disaggregation

During tissue disaggregation, an unwanted cell clumping can occur making a dispersion of single cells difficult.Since dissociation of tissue and isolation of single cells always is accompanied by rupture and lysis of some cells,DNA is released from these cells into the culture and dissociation medium respectively. Such a DNA content withinthe medium is described to be responsible for unwanted cell clumping during tissue disaggregation procedures. This effect can be avoided by adding deoxyribonuclease (DNase) to the dissociation medium and has been described for a variety of different cell and tissue types.

3.Reconstitute in sterile, double-distilled water to a final concentration of 10 mg/ml.Filtered through 0.2 ¦Ìm pore size membrane after  reconstitute.

4.For reconstitution of lyophilizate:

- Sterile, double-distilled water

- PBS (phosphate buffered saline)

- HBSS (Hank¡¯s balanced salt solution)

- Culture medium

Storage

The powder will be stable for 12 months under -20¡æ.DNase I retains activity for at least three years when stored at ¨C20 ¡ãC. Solutions of DNase I (10 mg/ml) in 0.15 M NaCl may lose <10% of its activity stored for a week in aliquots at ¨C20 ¡ãC. The same solutions stored in aliquots at 2¨C8 ¡ãC can lose ~20% activity. DNase I remains active in solution between pH 5 and 7 up to 60 ¡ãC for at least five hours. A 1 mg/ml solution in acetate buffer (pH 5.0) or Tris buffer (pH 7.2) loses activity at the rate of 6%/hour. At 68 ¡ãC DNaseI loses activity in <10 minutes.

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