D5025

Deoxyribonuclease I from bovine pancreas

Type IV, lyophilized powder, ≥2,000 Kunitz 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
• 유형: Type IV
• 양식: lyophilized powder
• 특이 활성도: ≥2,000 Kunitz units/mg protein
• 분자량: ~31 kDa
• 정제법: chromatography
• 구성: Protein, ≥80%
• 기술: DNA purification: suitable
• solubility: 0.15 M NaCl: soluble 5.0 mg/mL, clear, colorless
• 적합성: suitable for molecular biology

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 processing of rat brain tissue. This study showed that axonal growth on astrocytes is not inhibited by oligodendrocytes.[1] In another study, thawed fixed samples of E. coli were digested with DNAse I from Sigma along with other enzymes. The digestion was done before permeabilization and staining of the nucleic acids.[2]

Deoxyribonuclease I from bovine pancreas has been used in a study to investigate a two-dimensional zymogram analysis of nucleases in Bacillus subtilis. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effects of minor and major groove-binding drugs and intercalators on the DNA association of minor groove-binding proteins RecA and deoxyribonuclease I.

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⁺.[3] 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.[4] 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. Only minor amounts of D are found.[5] 2-Mercaptoethanol, chelators, sodium dodecyl sulfate (SDS)[6] and actin[7] are known to inhibit the enzyme activity.

단위 정의

One Kunitz unit will produce a ΔA₂₆₀ of 0.001 per min per mL at pH 5.0 at 25 °C, using DNA, Type I or III as substrate.

물리적 형태

Lyophilized powder containing calcium chloride

제조 메모

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