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
• 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

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.

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.[1]

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

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.

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] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[3] and actin[4] are known to inhibit the enzyme activity.

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
• wgk: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable