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D4263

Sigma-Aldrich

Deoxyribonuclease I from bovine pancreas

Standardized vial containing 2,000 Kunitz units of DNase I (D4527), vial of ≥0.25 mg total protein

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Synonym(s):
DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

Quality Level

form

powder

mol wt

~31 kDa

purified by

chromatography

packaging

vial of ≥0.25 mg total protein

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

application(s)

diagnostic assay manufacturing
diagnostic assay manufacturing

storage temp.

−20°C

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This Item
D4513D4527D5025
form

powder

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear(lit.)

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear

solubility

0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

mol wt

~31 kDa

mol wt

~31 kDa

mol wt

~31 kDa

mol wt

~31 kDa

Quality Level

200

Quality Level

300

Quality Level

300

Quality Level

300

Application

DNAse I is used to nick DNA as a first step to incorporate labeled bases into DNA. DNAse I from Sigma has been used along with other enzymes for tumor harvest and dissociation, during the isolation and molecular characterization of cancer stem cells in MMTV-Wnt-1 murine breast tumors.
Deoxyribonuclease I from bovine pancreas has been used in a study to determine that mammalian deoxyribonucleases I are classified into three types based on differences in their tissue concentrations. Deoxyribonuclease I from bovine pancreas has also been used in a study to compare the three primary structures of deoxyribonuclease isolated from bovine, ovine, and porcine pancreas.
Used for the removal of DNA from protein samples.

Biochem/physiol Actions

DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5′-phosphates. In the presence of Mg2+, DNAse I cleaves each strand of DNA independently and the cleavage sites are random. Both DNA strands are cleaved at approximately the same site in the presence of Mn2+. The pH optimum is found to be between 7 and 8. Divalent cations such as Mn2+, Ca2+, Co2+, and Zn2+ are activators of the enzyme. A concentration of 5 mM Ca2+ stabilizes the enzyme against proteolytic digestion. DNAse I from bovine pancreas consists of four chromatographically distinguishable components, A, B, C, and D, with molar ratios being 4:1:1 respectively. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Features and Benefits

Efficient DNA removal: Deoxyribonuclease I from bovine pancreas is an effective endonuclease that cleaves both strands of DNA randomly, making it a reliable choice for DNA removal from protein samples.

Versatile applications: This enzyme can be used to nick DNA, incorporate labeled bases into DNA, and isolate and molecularly characterize cancer stem cells in murine breast tumors. It is also useful in comparative studies of deoxyribonuclease isolated from bovine, ovine, and porcine pancreas.

Stable: The enzyme remains active for up to five hours at 60 °C between pH 5 and 7, and can be stored for up to a week at −20 °C with minimal loss of activity. This stability makes it a cost-effective and convenient option.

Unit Definition

One Kunitz unit will produce a ΔA260 of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III as substrate.

Preparation Note

The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 10 mg/mL solution of DNAse I in 0.15 M NaCl may lose <10% of its activity when stored for a week in aliquots at −20 °C. The same solutions stored in aliquots at 2-8 °C can lose approximately 20% activity. It remains active for up to five hours at 60 °C between pH 5 and 7, and loses activity in <10 minutes at 68 °C. It loses activity at the rate of 6%/hour in acetate buffer (pH 5.0) and tris buffer ((pH 7.2) at 1 mg/mL concentration.

Analysis Note

Protein determined by biuret.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Sambrook, J., and Russell, D.W
Molecular Cloning: A Laboratory Manual, 2(2), 5-5 null
Robert W Cho et al.
Stem cells (Dayton, Ohio), 26(2), 364-371 (2007-11-03)
In human breast cancers, a phenotypically distinct minority population of tumorigenic (TG) cancer cells (sometimes referred to as cancer stem cells) drives tumor growth when transplanted into immunodeficient mice. Our objective was to identify a mouse model of breast cancer
B J Gargiulo et al.
European cells & materials, 3, 9-18 (2003-10-17)
Chondrocytes undergo phenotypic alterations following extended periods in monolayer culture, i.e., they become bipolar and flattened, proliferate, and synthesise type I as opposed to type II collagen. This process has been termed chondrocyte dedifferentiation. Bistratene A is a macrolide polyether
S S Majumdar et al.
Biology of reproduction, 58(3), 633-640 (1998-03-25)
The purpose of the present study was to establish culture conditions for the in vitro study of the rhesus monkey Sertoli cell (Sc) at three major stages of development, namely infancy, adulthood, and the intervening prepubertal period. Conditions for the
R K Singh et al.
The American journal of pathology, 145(2), 365-374 (1994-08-01)
We investigated the influence of organ microenvironment on the angiogenic phenotype in human renal cell carcinoma (HRCC) cells. HRCC line SN12C was established in vitro from a surgical specimen, and metastatic line SN12PM6 was isolated from a lung metastasis produced

Protocols

To standardize a procedure for the enzymatic assay of Deoxyribonuclease I.

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