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Merck

D5025

Sigma-Aldrich

Desoxirribonucleasa I from bovine pancreas

Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein

Sinónimos:

ADNasa I, Desoxirribonucleato 5′-oligonucleótido-hidrolasa

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About This Item

Número de CAS:
Comisión internacional de enzimas:
Número CE:
Número MDL:
Código UNSPSC:
12352204
NACRES:
NA.54
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origen biológico

bovine pancreas

Nivel de calidad

tpo

Type IV

Formulario

lyophilized powder

actividad específica

≥2,000 Kunitz units/mg protein

mol peso

~31 kDa

purificado por

chromatography

composición

Protein, ≥80%

técnicas

DNA purification: suitable

solubilidad

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

idoneidad

suitable for molecular biology

aplicaciones

diagnostic assay manufacturing
diagnostic assay manufacturing

actividad extraña

Chymotrypsin ≤0.5%
Protease ≤0.05%
RNase ≤0.02%

Condiciones de envío

wet ice

temp. de almacenamiento

−20°C

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Aplicación

Utilizado para eliminación del ADN de las muestras proteicas.
DNAse I is used to nick DNA as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used in the processing of rat brain tissue. This study showed that axonal growth on astrocytes is not inhibited by oligodendrocytes.[1] In another study, thawed fixed samples of E. coli were digested with DNAse I from Sigma along with other enzymes. The digestion was done before permeabilization and staining of the nucleic acids.[2]
Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a two-dimensional zymogram analysis of nucleases in Bacillus subtilis. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effects of minor and major groove-binding drugs and intercalators on the DNA association of minor groove-binding proteins RecA and deoxyribonuclease I.

Acciones bioquímicas o fisiológicas

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+.[3] 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.[4] 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 their molar ratios being 4:1:1. Only minor amounts of D are found.[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[6] and actin[7] are known to inhibit the enzyme activity.

Definición de unidad

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.

Forma física

Lyophilized powder containing calcium chloride

Nota de preparación

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.

Nota de análisis

Protein determined by biuret.

Pictogramas

Health hazard

Palabra de señalización

Danger

Frases de peligro

Consejos de prudencia

Clasificaciones de peligro

Resp. Sens. 1

Código de clase de almacenamiento

11 - Combustible Solids

Clase de riesgo para el agua (WGK)

WGK 3

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable


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T Guindulain et al.
Applied and environmental microbiology, 63(11), 4608-4611 (1997-11-15)
Three nucleic acid dyes (SYTO-13, TOTO-1, and YOYO-1) were tested on cultures of Escherichia coli and marine prokaryote populations. These dyes stain the RNA and DNA in E. coli but only respond to DNA in marine populations, according to the
J W Fawcett et al.
Journal of cell science, 103 ( Pt 2), 571-579 (1992-10-01)
Axon growth in vitro may be inhibited by contact with oligodendrocytes, but most axons grow readily on the surface of astrocyte monolayers. Since both cell types are in close contact with one another in the damaged nervous system, we have
Enzymes of Molecular Biology
Weir, A. F.
Methods in Molecular Biology, 16 (1993)
Chii J Chan et al.
Biophysical journal, 112(6), 1063-1076 (2017-03-30)
Understanding the physical mechanisms governing nuclear mechanics is important as it can impact gene expression and development. However, how cell nuclei respond to external cues such as heat is not well understood. Here, we studied the material properties of isolated
T Liao
The Journal of biological chemistry, 250(10), 3831-3836 (1975-05-25)
In the course of experiments on the role of the COOH-terminal residues in pancreatic deoxyribonuclease, we undertook to ascertain whether the presence of sodium dodecyl sulfate would render the normally unavailable terminus susceptible to hydrolysis by carboxypeptidase A. When DNase

Protocolos

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

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

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

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

Questions

1–8 of 8 Questions  
  1. Good afternoon, if the stock concentration of 10mg/ml may loose 10% of its activity after one week, I was wondering if it is okey regarding stability to keep 500ug/ml aliquots at -20ºC for several weeks.

    1 answer
    1. As cited in the product listing and datasheet, this product in solution shows an approximate loss of activity of 10% when stored in aliquots at −20 °C for 1 week. Studies at lower dilutions have not been performed. Typically, solutions stored at higher concentrations are the most stable.

      Helpful?

  2. Is Kunitz unit same as unit here? I have seen some papers and they have U/ml for DNase. How can i conver it?

    1 answer
    1. Product D5025, Deoxyribonuclease I is given in Kunitz units. This product is a powder. When reconstituting the product with a solution, for example 15,000 Kunitz units in 1 ml of 0.15 M NaCl the end-user will have 15,000 Kunitz units/ml.

      Helpful?

  3. Are stock solutions of Product D5025, Deoxyribonuclease I (DNase I), stable?

    1 answer
    1. Solutions of DNAse I (10 mg/ml) 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. lose approximately 20% activity.

      Helpful?

  4. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  5. How should I prepare a solution of Product D5025, DNAse I?

    1 answer
    1. This enzyme can be reconstituted in 0.15 M NaCl at a concentration of 10 mg/ml.

      Helpful?

  6. How can I convert units of Product D5025, Deoxyribonuclease I (DNase I), to mass?

    1 answer
    1. The mg of solid for each unit package size will be indicated on the label of the package. For a particular lot, you can calculate the specific activity in units per mg solid by multiplying the specific activity in units per mg protein by the percent protein divided by 100.

      Helpful?

  7. How do Deoxyribonuclease I (DNase I), products D5025 and D4527  differ from each other?

    1 answer
    1. Both of these products have the same specific activity (>2000 units/mg protein) for Dnase I. The enzyme impurity levels for each, however, are different. D5025 has the following impurity specification: Chymotrypsin (<0.5%), Protease (<0.05%), and RNase (<0.02%). The impurity specifications for D4527 are: Chymotrypsin (<0.01%), Protease (<0.005%), and RNase (<0.01%).

      Helpful?

  8. What concentration of Product D5025, Deoxyribonuclease I (DNase I),  is required to remove DNA from solutions?

    1 answer
    1. DNA can be removed from preparations by incubation with 20 - 50 ug DNAse I in the presence of 50 mM Tris-HCl buffer, pH 7.5, 10 mM magnesium chloride, at 37°C. for 60 min.

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