생물학적 소스
bovine pancreas
Quality Level
유형
Type II
양식
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
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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.
신호어
Danger
유해 및 위험 성명서
예방조치 성명서
Hazard Classifications
Resp. Sens. 1
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
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