D2821

Desoxyribonuklease I Rind

recombinant, expressed in Pichia pastoris, lyophilized powder, RNAse and protease, free

• Synonym(e): DNAse I, Desoxyribonukleat-5′-oligo-nukleotidhydrolase
• UNSPSC Code: 12352204
• NACRES: NA.54
• EG-Nummer: 3.1.21.1 (BRENDA, IUBMB)
• MDL number: MFCD00130918
• Specific activity: ≥4,000 units/mg protein
• Biological source: bovine
• Recombinant: expressed in Pichia pastoris

Eigenschaften

• biological source: bovine
• Quality Segment: 300
• recombinant: expressed in Pichia pastoris
• form: lyophilized powder
• specific activity: ≥4,000 units/mg protein
• mol wt: ~39 kDa
• technique(s): DNA extraction: suitable
• input: general use sample(s)
• solubility: H₂O: soluble (pH 4.0-9.0)
• suitability: suitable for molecular biology
• application(s): diagnostic assay manufacturing
• foreign activity: RNAse and protease, free
• storage temp.: 2-8°C

Beschreibung

Application

The enzyme from Sigma has been used for the digestion of DNA extracted from Agave spp. clones. The enzyme was used during a study that investigated the effect of epigenetic changes on the regulatory expression of KNOTTED1-like HOMEOBOX (KNOX) transcription factors.

Wird verwendet für das Entfernen von DNA aus Proteinproben.

Deoxyribonuclease I bovine has been used in a study to investigate the inhibition of DNA polymerase by extracts of rat liver. Deoxyribonuclease I bovine has also been used in a study to investigate the effects of ionic strength on enzymic activity.

Deoxyribonuclease I bovine has been used in the preparation of cold cell lysis buffer, complete RNA lysis buffer, cell lysis buffer for testing the expression of recombinant tagged protein.

Biochem/physiol Actions

DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5′-phosphates. 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. A concentration of 5 mM Ca²⁺ stabilizes the enzyme against proteolytic digestion. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

Erzeugt aus ein- und doppelsträngiger DNA mittels Verdau eine Mischung aus Mono- und Oligonukleotiden, die 5′-Phosphate und 3′-OH-Termini besitzen. Diese katalytische Aktivität ist divalent ionenabhängig. In Gegenwart von Mg²⁺ hydrolysiert DNase I jeden Strang von Doppelstrang-DNA unabhängig und nach dem Zufallsprinzip. In Gegenwart von Mn²⁺ können beide Stränge aufgespalten werden.

Features and Benefits

• RNA-Aufreinigung durch Entfernen von DNA
• Präparation von DNA für die Nick-Translation¹
• Footprinting-Assays für die Bestimmung von DNA-Protein-Interaktionen²

Physical form

supplied as a lyophilized powder containing glycine as a stabilizer

Preparation Note

Produced without using any animal cells or animal derived materials.

The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 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 upto five hours at 60 °C 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.

Other Notes

One unit will produce a ΔA₂₆₀ of 0.001 per min per mL reaction mixture using calf thymus DNA at pH 5.0 and 25°C

Sicherheitshinweise

• Lagerklasse: 11 - Combustible Solids
• Was ist los?: WGK 3
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable