DNase I, Bovine pancreas

DNase I (EC 3.1.21.1) is an enzyme that degrade DNA, it plays a key role in the cleavage of extracellular DNA is crucial for limiting the inflammatory response and maintaining homeostasis. Exogenous deoxyribonuclease shows beneficial effects in inflammatory diseases and cancer^[1]. In Vitro: Product Information
The activity of DNase Ⅰ is dependent on Ca²⁺ and can be activated by divalent metal ions such as Co²⁺, Mn²⁺, Zn²⁺, and others.
In the presence of Mg²⁺, the enzyme can randomly recognize and cleave any site on either strand of DNA; whereas, in the presence of Mn²⁺, it can simultaneously recognize both strands of DNA and cleave at nearly identical sites. This product is extracted from bovine pancreas and operates optimally within a pH range of 7-8. It has a molecular weight of approximately 31 kDa and an isoelectric point of around 6.0.

Instructions
Inactivation or Inhibition of DNase I:
Dissolved DNase I can be inactivated by heating at 65°C for 10 min. Phenol-chloroform extraction can also inactivate DNase I. Metal ion chelators, zinc ions at millimolar concentrations, 0.1% SDS, reducing agents such as DTT and β-mercaptoethanol, and salt concentrations above 50-100 mM all significantly inhibit DNase I.
Usage Method for Protein Extraction Experiments (for reference only):
1)Preparation of storage solution: 5-10 mg/mL dissolved in 0.15 M NaCl, stored in the refrigerator, it is recommended to be used within 1 week.
2)Reaction System: Add DNase I stock solution to the protein extraction buffer at a 1/100 volume ratio (to achieve a final concentration of 20 U/mL) and 1 M MgCl2 at a 1/100 volume ratio.
3)Reaction Conditions: Incubate at 37°C for 30-60 min. Proceed with subsequent protein extraction experiments.

Notes
1.Once the solution is prepared, please store it in aliquots to avoid product failure caused by repeated freezing and thawing.
2.Since EDTA can chelate Ca²⁺ and Mg²⁺ required for enzyme activity, EDTA should be removed from the initial protein lysis buffer, as it will reduce the digestive capacity of DNase I.
In Vivo: Deoxyribonuclease (0.1 U; i.p.; once daily for 3 days) inhibits liver metastasis, besides results in a greater prolongation of the survival period by combining surgical removal of the primary tumour mass^[1].