D4527

Deoxyribonuclease I from bovine pancreas

Type II, lyophilized powder, Protein ≥80 %, ≥2,000 units/mg protein

• 동의어(들): DNase I, Deoxyribonucleate 5′-oligonucleotido-hydrolase
• CAS Number: 9003-98-9
• 효소 위원회 번호: 3.1.21.1 (BRENDA, IUBMB)
• EC Number: 232-667-0
• MDL number: MFCD00130918
• UNSPSC 코드: 12352204
• NACRES: NA.54

속성

• 생물학적 소스: bovine pancreas
• Quality Level: 300
• 유형: Type II
• 형태: lyophilized powder
• 특이 활성도: ≥2,000 units/mg protein
• 분자량: ~31 kDa
• 정제법: chromatography
• 구성: Protein, ≥80%
• 기술: DNA extraction: suitable
• solubility: 0.15 M NaCl: soluble 5.0 mg/mL, clear
• 적합성: suitable for molecular biology
• 응용 분야: diagnostic assay manufacturing; diagnostic assay manufacturing
• 외래 활성: Chymotrypsin ≤0.01%; Protease ≤0.005%; RNase ≤0.01%
• 배송 상태: wet ice
• 저장 온도: −20°C

설명

애플리케이션

DNAse I is used to nick DNA, as a first step to incorporate labeled bases into DNA. The enzyme from Sigma has been used in the nuclease stock along with RNAse during cell lysate preparation of Madin-Darby canine kidney (MDCK) II cell lines. It has also been used during RNA extraction from Sindbis virus.

Deoxyribonuclease I from bovine pancreas has been used for the identification, localization and expression of two novel human genes similar to deoxyribonuclease I. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate a new approach to obtaining high-activity RNase, DNase, cholesterolesterase, and trypsin from cattle pancreas.

Used for the removal of DNA from protein samples.

생화학적/생리학적 작용

DNase I is an endonuclease that acts on phosphodiester bonds adjacent to pyrimidines to produce polynucleotides with terminal 5′-phosphates. In the presence of Mg²⁺, DNAse I cleaves each strand of DNA independently and the cleavage sites are random. Both DNA strands are cleaved at approximately the same site in the presence of Mn²⁺. 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. DNAse I from bovine pancreas consists of four chromatographically distinguishable components, A, B, C, and D, with their molar ratios being 4:1:1 respectively. Only minor amounts of D are found. 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS) and actin are known to inhibit the enzyme activity.

특징 및 장점

Effective for nicking DNA and removal of DNA from protein samples

단위 정의

One Kunitz unit will produce a change in A₂₆₀ of 0.001 per minute per ml at pH 5.0 at 25 °C using DNA, Type I or III, as the substrate.

물리적 형태

Contains calcium chloride

제조 메모

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 up to five hours at 60 °C between pH 5 and 7, 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.

분석 메모

Protein determined by biuret.

안전성 정보

• 픽토그램: GHS08
• 신호어: Danger
• 유해 및 위험 성명서: H334
• 예방조치 성명서: P261 - P284 - P501
• Hazard Classifications: Resp. Sens. 1
• Storage Class Code: 11 - Combustible Solids
• WGK: WGK 3
• Flash Point (°F): Not applicable
• Flash Point (°C): Not applicable