| 规格或纯度 | ≥2,000 Kunitz units/ml |
|---|---|
| 英文名称 | Deoxyribonuclease I from Bovine Pancreas(RNase & Protease Free,Solution) |
| 别名 | 脱氧核糖核酸酶 I |
| 英文别名 | Deoxyribonucleate 5′-oligonucleotido-hydrolase;DNase I |
| 单位定义 | 1 Unit causes an increase in absorbance at 260nm of 0.001 per minute per ml at 25°C when acting upon highly polymerized DNA at pH 5.0. Note: Kunitz units as reported by other suppliers can be 2 to 4 times higher than Kunitz units as measured at Aladdin. A |
| 生化机理 | 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+. 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. The pH optimum is found to be between 7 and 8. 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)6 and actin7 are known to inhibit the enzyme activity. |
| 储存温度 | -20°C储存 |
| 运输条件 | 超低温冰袋运输 |
| 产品介绍 |
Bovine pancreatic deoxyribonuclease is an endonuclease which splits phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide yielding polynucleotides with free hydroxyl group at the 3' position and phosphate group at the 5' position. The average chain length of a limit digest is a tetranucleotide.用于从蛋白质样品中除去 DNA。 Deoxyribonuclease I from bovine pancreas has been used in a study to compare several procedures for reducing RNase contamination in preparations of DNase. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effect of the composition of sodium dodecyl sulfate preparations on the renaturation of enzymes after polyacrylamide gel electrophoresis. BbpDNase I?is not base nor sequence specific; however, it does not cleave randomly. It shows preference for cleavage at the 5’ side of pyrimidines, and is particularly pronounced in alternative copolymers (Bernardi et al. 1975, and Lomonossoff et al. 1981). It has been shown that variations in the twist angle are recognized by DNase I?(Dickerson and Drew 1981). The specificity of DNase I?also depends on the divalent cations present. In the presence of Ca2+ and Mg2+, it causes single strand breaks, and in the presence of Mn2+ double strand breaks have been reported (Junowicz and Spencer 1973, and Campbell and Jackson 1980). Protein Accession Number: P00639CATH Classification (v. 3.2.0): Class:?Alpha Beta Architecture:?4-Layer Sandwich Topology:?Deoxyribonuclease I; Chain A? Molecular Weight:?29.1 kDa (Theoretical) Optimal pH:?7.8 Isoelectric Point:?5.08 (Theoretical) Extinction Coefficient:? 36,750 cm-1 M-1 E1%,280 =?11.1 Active Site Residues: Tyrosine (Y87) Glutamate (E100) Histidine (H156) Activators: Bivalent metal ions (Junowicz and Spencer 1973b, Poulos and Price 1972, Price 1969, and Price 1975) Inhibitors: 2-mercaptoethanol (Laskowski 1971)? 2-nitro-5-thiocyanobenzoic acid (Moore 1981) Actin (Mannherz et al. 2008, and Lazarides and Lindberg 1974) Alfatoxin B2a, G2, G2a, and M1 (non-competitive) (Schabort 1970) EGTA (Moore 1981) and EDTA (Junowicz and Spencer 1973) Sodium dodecyl sulfate (Liao 1975a) Calf spleen inhibitor protein (Laskowski 1971) Carbodiimide and cholesterol sulfate (Iwamori 2000) Iodoacetate (Moore 1981) ?Applications: DNA removal in primary cell isolation: decreases viscosity providing better yields DNA removal in bioprocessing applications Removing genomic DNA from RNA preparations prior to RT-PCR in vitro transcription Nick translation DNase footprinting Actin binding UV crosslinking of proteins to nucleic acids Radioactive labeling |
| EC号 | 232-667-0 |
|---|---|
| 酶学委员会编号 | 3.1.21.1 |
| 分子式 |
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| PubChem CID | 597667 |
| WGK Germany | 3 |
|---|---|
| RTECS | RF0750000 |
| 个人防护装备 | Eyeshields, Gloves, type N95 (US), type P1 (EN143) respirator filter |
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