D2821

Deoxyribonuclease I bovine

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

• Sinónimos: DNAse I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• Comisión internacional de enzimas: 3.1.21.1 (BRENDA, IUBMB)
• MDL number: MFCD00130918
• UNSPSC Code: 12352204
• NACRES: NA.54

Propiedades

• biological source: bovine
• Quality Level: 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

Descripción

Application

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,[1] complete RNA lysis buffer,[2] cell lysis buffer for testing the expression of recombinant tagged protein.[3]

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.[4]

Used for the removal of DNA from protein samples.

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.[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[6] and actin[7] are known to inhibit the enzyme activity.

Digests single- and double-stranded DNA to a mixture of mono- and oligonucleotides carrying 5′ phosphates and 3′ OH termini. This catalytic activity is divalent ion-dependent. In the presence of Mg²⁺, DNase I hydrolyzes each strand of double-stranded DNA randomly and independently. In the presence of Mn²⁺, both strands can be cleaved.

Features and Benefits

• RNA purification by removing DNA
• Prepare DNA for nick translation¹
• Footprinting assays to determine DNA-protein interactions²

Unit Definition

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

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.

Información de seguridad

• Storage Class: 11 - Combustible Solids
• wgk_germany: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable