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| Product Name | DNase I, from Bovine Pancreas (Powder) |
| Catalog No. | IVDRM-0195 |
| Description | Deoxyribonuclease I (DNase I) is an endonuclease present in a wide variety of cells and tissues. It cleaves phosphodiester bonds adjacent to pyrimidines, generating polynucleotides with 5'-phosphate and 3'-hydroxyl termini, with tetranucleotides as the smallest average digestion products. DNase I can act on many forms of DNA, including single-stranded DNA, double-stranded DNA, and even chromatin, whose cleavage rate is affected by histones. Its activity is Ca2+-dependent and can be activated by divalent metal ions such as Co2+, Mn2+, and Zn2+; 5 mM Ca2+ protects the enzyme from hydrolysis. In the presence of Mg2+, the enzyme randomly recognizes and cleaves any site on either DNA strand, whereas in the presence of Mn2+ it recognizes both strands simultaneously and cuts at nearly identical positions. DNase I was first isolated from pancreas, and mammalian pancreas remains one of the main sources of this enzyme. This product is purified from bovine pancreas and is commonly used in molecular biology to remove DNA from protein preparations or to introduce nicks into DNA for the incorporation of labeled bases. It is supplied as a powder with an activity of at least 2000 Kunitz Units per mg of protein. |
| Intended Use | Commonly used in molecular biology to remove DNA from protein preparations, or to introduce nicks into DNA so that labeled bases can be incorporated. |
| Principle / Technology | DNase I is an endonuclease that cleaves phosphodiester bonds adjacent to pyrimidines, producing polynucleotides with 5'-phosphate and 3'-hydroxyl ends. Its activity depends on Ca2+ and is activated by divalent metal ions. With Mg2+, it randomly cleaves either DNA strand; with Mn2+, it cleaves both strands at nearly the same positions. |
| Sample Type | Single-stranded DNA, double-stranded DNA, chromatin-containing samples, and protein preparations contaminated with DNA. |
| Performance Range / Specifications | CAS NO.: 9003-98-9 Molecular weight: ~31 kDa Activity: >=2000 Kunitz Units/mg protein Type: IV Optimal pH: 7-8 Appearance: white to light yellow powder Purity: Protein >=80% by Biuret |
| Specificity | Targets phosphodiester bonds adjacent to pyrimidines in DNA; the smallest average digestion products are tetranucleotides. |
| Reaction Conditions / Protocol | Optimal working pH is 7-8. Activity is Ca2+-dependent and is activated by divalent metal ions such as Co2+, Mn2+, and Zn2+; 5 mM Ca2+ protects the enzyme from hydrolysis. In the presence of Mg2+, the enzyme randomly cleaves either DNA strand; in the presence of Mn2+, it cleaves both strands at nearly identical sites. |
| Components / Formulation | Supplied as a powder. |
| Storage Conditions | Store the lyophilized powder at -20°C. |
| Shelf Life | 2 years when stored at -20°C. |
| Package Specifications | 25 mg; 100 mg; 1 g |
| Product Form | Powder |
| Quality Control | Activity verified at >=2000 Kunitz Units/mg protein; protein content >=80% by Biuret. |
| Purity | Protein: >=80% by Biuret |
| Activity / Unit Definition | >=2000 Kunitz Units/mg protein. One Kunitz unit is defined as the amount of enzyme that causes an increase in absorbance at 260 nm (delta A260) of 0.001 per mL per minute when acting on a DNA substrate at 25°C and pH 5.0. |
| Molecular Weight | ~31 kDa |
| Source / Origin | Bovine pancreas |
| pH Range / Optimal pH | Optimal pH 7-8 |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Valid for 2 years when the powder is stored at -20°C. |
| Compatibility | Activated by divalent metal ions (Mg2+, Mn2+, Ca2+, Co2+, Zn2+); inhibited by beta-mercaptoethanol, chelating agents, SDS, and actin. |
| Recommended Buffer System | Dissolve in an appropriate buffer containing the required divalent metal ions. Dissolving in water is not recommended, as the enzyme is unstable in water and lacks the ionic activation needed for optimal activity. |
| Application Notes / Precautions | Kunitz Units and standard units (U) are different measures of activity and cannot be directly converted. To calculate the mass needed for a given number of Kunitz Units, use the lot-specific activity on the COA. For example, for a lot assayed at 2500 Kunitz Units/mg, 150 Kunitz Units corresponds to 150/2500 = 0.06 mg. Dissolving the powder in water is not recommended; water lacks the activating ions required for optimal enzyme activity and results in poor stability. A calcium chloride-containing lyophilized format of the same enzyme is also available; both formats share the same application of removing DNA from protein preparations. |
For research use only, not for clinical use.
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