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| Product Name | DNase I, from Bovine Pancreas (15 KU) |
| Catalog No. | IVDRM-0194 |
| Description | Deoxyribonuclease I (DNase I) is an endonuclease found in a wide variety of cells and tissues. It cleaves phosphodiester bonds adjacent to pyrimidines, yielding polynucleotides with 5'-phosphate and 3'-hydroxyl termini, with tetranucleotides as the smallest average digestion products. DNase I acts on many forms of DNA, including single-stranded DNA, double-stranded DNA, and even chromatin, although its cleavage rate on chromatin is influenced by histones. Its activity is Ca2+-dependent and can be stimulated by divalent metal ions such as Co2+, Mn2+, and Zn2+; 5 mM Ca2+ protects the enzyme against hydrolysis. In the presence of Mg2+, the enzyme randomly recognizes and cleaves sites on either DNA strand, whereas in the presence of Mn2+ it cuts both strands at nearly identical sites. DNase I was first isolated from pancreas, and mammalian pancreas remains one of its principal sources. This product is purified from bovine pancreas and consists of four chromatographically resolved components (A, B, C, and D) in a molar ratio of 4:1:1, with component D present in very small amounts. In molecular biology, it is commonly used to remove DNA from protein preparations or to introduce nicks into DNA for the incorporation of labeled bases. It is supplied as a lyophilized powder containing calcium chloride, 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 strand of DNA; 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 brown lyophilized powder Purity: Protein >=80% by Biuret Solubility: 5 mg/mL in 0.15 M NaCl, yielding a colorless, clear solution |
| 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. Reconstituted stock solutions are best used immediately or prepared fresh; store at -20°C. |
| Components / Formulation | Lyophilized powder containing calcium chloride. |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 2 years when stored at -25 to -15°C. |
| Package Specifications | 15 KU; 10x15 KU |
| Product Form | Lyophilized powder (contains calcium chloride) |
| 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 |
| Expiration Date / Stability | Valid for 2 years when stored at -25 to -15°C. Reconstituted solutions should be used promptly and 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 0.15 M NaCl (soluble to 5 mg/mL, yielding a colorless, clear solution). |
| Application Notes / Precautions | The KU specification can be converted to mass using the lot-specific Kunitz activity. For example, the 15 KU size corresponds to approximately 7.5 mg (15,000 Kunitz Units / >=2000 Kunitz Units per mg protein). Reconstituted DNase I solutions are best used immediately or prepared fresh; store at -20°C. Primary applications: removal of DNA from protein preparations and introduction of nicks into DNA for labeled-base incorporation. |
For research use only, not for clinical use.
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