ROCHE DNase I recombinant, grade I, from bovine pancreas, expressed in Pichia pastoris

UNSPSC Code: 12352200

RIDADR: NONH for all modes of transport

Features and Benefits:
Contents
Lyophilizate

Application:
• DNase I, recombinant, grade I is used for:Eliminating DNA during protein isolation procedures
• Analysis of chromatin structure
• Eliminating DNA during sample preparation

General description:
Properties: Double-strand-specific endonuclease that requires bivalent cations for maximal activity.
Activity: 10,000U/bottle according to Kunitz (+25°C; DNA as substrate).
Unit definition: One unit is the enzyme activity that causes an increase in the absorbance of 0.001 per minutes under assay conditions.

Other Notes:
For life science research only. Not for use in diagnostic procedures.

Preparation Note:
Activator: The enzyme requires divalent cations for maximal activity. The specificity of the reaction depends on the nature of the cation used. In the presence of Mg²⁺ DNase I causes single-stranded nicks in dsDNA, while in the presence of Mn²⁺ the enzyme produces double-stranded breaks.
Storage conditions (working solution): Long-term Storage of the Dissolved Enzyme

The solvent generally recommended for DNase I is water. When DNase I is reconstituted in water, the solution can be kept for 2 days at 2 to 8 °C and for 1 month at -15 to -20 °C. For best results, prepare appropriate aliquots and avoid repeated freezing and thawing. However, for long-term storage, carefully dissolve DNase I in one of the buffers listed below. In buffer 1, the enzyme will be stable for several weeks. The enzyme will be stable for at least 18 months if it is dissolved in buffer 2, 3, or 4 and stored at the recommended temperatures.

Information note: Do not vortex the enzyme while dissolving! Make sure the solution contains at least 1 mg enzyme/ml.

Buffer 1: 20 mM Tris-HCl, 20 mM MgCl₂, 5 mM CaCl₂, 0.1 mM dithioerythritol, 0.1 mM EDTA, 50% (v/v) glycerol, pH 8. Store this solution at -15 to-25 °C; it will not freeze at this temperature and will be stable for several weeks.
Buffer 2: 20 mM Tris-HCl, 50 mM NaCl, 1 mM dithioerythritol, 100 μg/mL BSA, 50% glycerol (v/v), pH 7.6. Store this solution at -15 to -25 °C for up to 18 months; it will not freeze at this temperature.
Buffer 3: 20 mM Tris-HCl, 1 mM MgCl₂, 50% (w/v) glycerol, pH 7.5. Store this solution at -15 to -25 °C for up to 18 months; it will not freeze at this temperature.
Buffer 4: 20 mM Tris-HCl, 1 mM MgCl₂, pH 7.5. Aliquot in appropriate amounts (e.g., 10 μl), freeze the aliquots quickly on dry ice, and store at -60 °C or below for up to 18 months. Thaw only the amount needed for each experiment. Do not refreeze. Discard any leftover thawed solution.