产品描述

脱氧核糖核酸酶I（DNase I），即Deoxyribonuclease I，一种发现于多种细胞和组织的核酸内切酶，靶向切割邻近嘧啶的磷酸二酯键，产生5’端为磷酸基团、3’端为羟基的多聚核苷酸，平均消化产物最小为多聚四核苷酸。DNase可催化多种形式DNA，如单链DNA、双链DNA，甚至染色质（其切割速率受组蛋白影响）。最佳的工作范围是pH7-8，DNase的活性依赖于Ca²⁺，并可被二价金属离子如Co ²⁺，Mn²⁺，Zn²⁺等激活。5 mM Ca²⁺可保护酶使其不被水解。在Mg²⁺存在下，该酶可随机识别和切断DNA任一条链上的任意位点；而在Mn²⁺存在的条件下，可同时识别DNA的两条链并在几乎相同的位点进行切割。DNase I最早从胰腺中分离而来，至今哺乳动物胰腺也是该酶的最主要的来源之一。
本品来自牛胰腺，由四种色谱区分的组分A，B，C，D构成，摩尔比为4:1:1，而D组分非常少。分子生物学实验中常用于清除蛋白中的DNA，或者用于向DNA中引入缺口使标记碱基插入DNA。本品以含氯化钙的冻干粉形式供应，酶活力≥2000 Kunitz Units/mg蛋白。

产品性质

运输和保存方法 

冰袋运输。冻干粉末于-20℃保存，有效期2年。

注意事项

1）为了您的安全和健康，请穿实验服并戴一次性手套操作。
2）本产品仅作科研用途！

DNase Ⅰ储存溶液

20 mM sodium acetate（pH 6.5），5 mM CaCl₂，0.1 mM PMSF，50%甘油。

DNase Ⅰ失活或抑制

加入EDTA至终浓度为2.5 mM后，65℃加热10 min可使DNase Ⅰ失活。酚氯仿抽提也可以使DNase I失活。金属离子螯合剂，达到毫摩尔/升浓度的锌离子，0.1%的SDS，DTT、巯基乙醇等还原剂，50-100 mM以上盐浓度均对DNase Ⅰ有显著抑制作用。

使用方法（应用于蛋白提取实验，仅供参考）

1）反应体系：蛋白提取液中按照1/100体积加入DNase I储存液（使其终浓度为20 U/mL），1/100体积加入1 M MgCl₂。

2）反应条件：37℃，30-60 min。继续后续蛋白提取实验即可。

【注】由于EDTA能螯合酶活性需要的Ca²⁺和Mg²⁺，蛋白初始裂解液中要去除EDTA，否则会降低DNase I的消化能力。

HB210811

 Q：DNase Ⅰ储存溶液配置后可保存多久？保存温度是多少？

A：最好快速用完，或者现配现用。-20℃保存。

Q：主要用途是什么？

A：常用于清除蛋白中的DNA，或者用于向DNA 中引入缺口使标记碱基插入DNA。

[1] Xu Y, Hu Y, Xu T, et al. RNF8-mediated regulation of Akt promotes lung cancer cell survival and resistance to DNA damage. Cell Rep. 2021;37(3):109854. doi:10.1016/j.celrep.2021.109854(IF:9.423)
[2] Jain S, Hu C, Kluza J, et al. Metabolic targeting of cancer by a ubiquinone uncompetitive inhibitor of mitochondrial complex I. Cell Chem Biol. 2022;29(3):436-450.e15. doi:10.1016/j.chembiol.2021.11.002(IF:8.116)
[3] Sun H, Chen D, Zhan S, et al. Design and Discovery of Natural Cyclopeptide Skeleton Based Programmed Death Ligand 1 Inhibitor as Immune Modulator for Cancer Therapy. J Med Chem. 2020;63(19):11286-11301. doi:10.1021/acs.jmedchem.0c01262(IF:6.205)
[4] Zhao X, Hu S, Zeng L, et al. Irradiation combined with PD-L1^-/- and autophagy inhibition enhances the antitumor effect of lung cancer via cGAS-STING-mediated T cell activation. iScience. 2022;25(8):104690. Published 2022 Jun 30. doi:10.1016/j.isci.2022.104690(IF:6.107)

DNase I is an endonuclease that can digest single-stranded and double-stranded DNA to produce single deoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides. It can hydrolyze the phosphodiester bond to produce monodeoxynucleotides and oligodeoxynucleotides containing 5'-phosphate groups and 3'-OH groups. The average digestion product is the smallest polytetranucleotide. DNase I can catalyze many forms of DNA, such as single-stranded DNA, double-stranded DNA, and even chromatin (its cutting rate is affected by histones).The optimum pH range is 7-8. The activity of DNase I depends on Ca²⁺ and can be activated by divalent metal ions, such as Co²⁺, Mn²⁺, Zn²⁺, etc. 5 mM Ca²⁺ can protect the enzyme from being hydrolyzed. In the presence of Mg²⁺, the enzyme can recognize and cut any site on any strand of DNA randomly; and in the presence of Mn²⁺, it can recognize two strands of DNA at the same time and cut at almost the same site to form blunt ends, Or sticky ends with 1-2 nucleotides protruding. DNase I is widely used in the preparation of DNA-free RNA; remove the template DNA after in vitro transcription; prepare DNA-free RNA before RT-PCR and RT-qPCR reactions; combine with DNA polymerase I to perform DNA labeling through nick translocation; DNA fragmentation library construction.

This product is produced in accordance with GMP process requirements, and the product is provided in liquid form.

Product Properties

Contents

Shipping and Storage

Deoxyribonuclease I (DNase I) GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

Experimental methods

Plasmid template digestion

1. Reaction system:

Use the RNase-free centrifuge tube and pipette tip to prepare the following reaction system:

【Note】1×DNase I Buffer: 10 mM Tris-HCl, 2.5 mM MgCl₂, 0.5 mM CaCl₂, pH7.6 @25℃

2. Reaction conditions

37℃, 15-30 min later, add a final concentration of 2.5 mM EDTA solution and mix well at 65℃ for 10 min. The processed template can be used in subsequent reactions such as capping reaction

DNase Ⅰ inactivation or inhibition

After adding EDTA to a final concentration of 2.5 mM, heating at 65°C for 10 min can inactivate DNase I. Phenol and chloroform extraction can also inactivate DNase I. The following conditions all have significant inhibitory effect on DNase I: Metal ion chelating agents, zinc ions with a concentration of millimoles/liter, 0.1% SDS, DTT, mercaptoethanol and other reducing agents,the salt concentrations above 50-100 mM.

Note

1. Enzymes should be stored in an ice box or on an ice bath when used, and should be stored at -20°C immediately after use.

2. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

HB230406

Q：是无菌的产品吗？

A：是的。

[1] Lu Z, Liu H, Song N, et al. METTL14 aggravates podocyte injury and glomerulopathy progression through N⁶-methyladenosine-dependent downregulating of Sirt1. Cell Death Dis. 2021;12(10):881. Published 2021 Sep 27. doi:10.1038/s41419-021-04156-y(IF:8.469)
[2] Yu Y, Wang Y, Zhang W, et al. Biomimetic periosteum-bone substitute composed of preosteoblast-derived matrix and hydrogel for large segmental bone defect repair. Acta Biomater. 2020;113:317-327. doi:10.1016/j.actbio.2020.06.030(IF:7.242)

产品描述

脱氧核糖核酸酶I（DNase I），即Deoxyribonuclease I，一种发现于多种细胞和组织的核酸酶，属核酸内切酶，靶向切割邻近嘧啶的磷酸二酯键，产生5’端为磷酸基团、3’端为羟基的多聚核苷酸，平均消化产物最小为多聚四核苷酸。DNase可催化多种形式DNA，如单链DNA、双链DNA，甚至染色质（其切割速率受组蛋白影响）。最佳的工作范围是pH7-8，DNase的活性依赖于Ca²⁺，并可被二价金属离子如Co²⁺，Mn²⁺，Zn²⁺等激活。5 mM Ca²⁺可保护酶使其不被水解。在Mg²⁺存在下，该酶可随机识别和切断DNA任一条链上的任意位点；而在Mn²⁺存在的条件下，可同时识别DNA的两条链并在几乎相同的位点进行切割。DNase I最早从胰腺中分离而来，至今哺乳动物胰腺也是该酶的最主要的来源之一。
本品来自牛胰腺，分子生物学实验中常用于清除蛋白中的DNA，或者用于向DNA中引入缺口使标记碱基插入DNA。本品以粉末形式供应，酶活力≥2000 Kunitz Units/mg蛋白。

产品性质

运输和保存方法

冰袋运输。冻干粉末于-20℃保存，有效期2年。

注意事项

1）为了您的安全和健康，请穿实验服并佩戴一次性手套操作。
2）本产品仅作科研用途！

DNase Ⅰ储存溶液

20 mM sodium acetate（pH 6.5），5 mM CaCl₂，0.1 mM PMSF，50%甘油。

DNase Ⅰ失活或抑制

加入EDTA至终浓度为2.5 mM后，65℃加热10 min可使DNase Ⅰ失活。酚氯仿抽提也可以使DNase I失活。金属离子螯合剂，达到毫摩尔/升浓度的锌离子，0.1%的SDS，DTT、巯基乙醇等还原剂，50-100 mM以上盐浓度均对DNase Ⅰ有显著抑制作用。

使用方法（应用于蛋白提取实验，仅供参考）

1）反应体系：蛋白提取液中按照1/100体积加入DNase I储存液（使其终浓度为20 U/mL），1/100体积加入1 M MgCl₂。

2）反应条件：37℃，30-60 min。继续后续蛋白提取实验即可。

【注】：由于EDTA能螯合酶活性需要的Ca²⁺和Mg²⁺，蛋白初始裂解液中要去除EDTA，否则会降低DNase I的消化能力。

HB210721

Q：10607ES 和 10608ES 的区别？

A：10607 以含氯化钙的冻干粉形式供应；10608 以粉末形式供应。两者用途一样，用于清除蛋白中的 DNA。

Q: 我想问一下这个Kunitz跟u如何换算呢？那如果说反应体系中需要加入150U，那我该怎么换算呀?

A: Kunitz Units和U是不同的单位，不能直接换算，它的酶活就是Kunitz Units不是U，是孔尼茨单位，不能换算的。

如果是要要加入150U是没法计算的，但如果是要150 Kunitz Units，可以根据每个批次的COA上的测试的酶活情况计算。比如，某批次酶活是2500Kunitz Units/mg，那就是加入150/2500=0.06mg。

[1] Wang Z, Gong X, Li J, et al. Oxygen-Delivering Polyfluorocarbon Nanovehicles Improve Tumor Oxygenation and Potentiate Photodynamic-Mediated Antitumor Immunity. ACS Nano. 2021;15(3):5405-5419. doi:10.1021/acsnano.1c00033(IF:15.881)
[2] Huang Y, Chen Y, Zhou S, et al. Dual-mechanism based CTLs infiltration enhancement initiated by Nano-sapper potentiates immunotherapy against immune-excluded tumors. Nat Commun. 2020;11(1):622. Published 2020 Jan 30. doi:10.1038/s41467-020-14425-7(IF:12.121)
[3] Wang Y, Gong X, Li J, et al. M2 macrophage microvesicle-inspired nanovehicles improve accessibility to cancer cells and cancer stem cells in tumors. J Nanobiotechnology. 2021;19(1):397. Published 2021 Nov 27. doi:10.1186/s12951-021-01143-5(IF:10.435)
[4] Liu J, Shen Z, Tang J, et al. Extracellular DNA released by glycine-auxotrophic Staphylococcus epidermidis small colony variant facilitates catheter-related infections. Commun Biol. 2021;4(1):904. Published 2021 Jul 22. doi:10.1038/s42003-021-02423-4(IF:6.268)