全能核酸酶（UltraNuclease），又称非限制性核酸内切酶、广谱核酸酶；是一种来源于 Serratia Marcescen 的非特异性核酸内切酶，可在链内任意核苷酸间进行切割，将核酸完全消化成2-5个碱基长度的5'-单磷酸寡核苷酸，能够在非常广泛的条件下降解各种形式的 DNA 和 RNA，广泛用于去除生物制品中的核酸。
【反应条件 Reaction conditions】
| 条件 Condition | 最佳 Optimum | 有效 Effective | | Mg2+ | 1-2 mM | 1-10 mM | | pH | 8-9 | 6-10
| | 温度/ Temperature | 37°C | 0-42°C | | DTT | 0-100 mM | >0 mM | | 巯基乙醇/ Mercaptoethanol | 0-100 mM | >0 mM | | 单价阳离子/ Monovalent cation | 0-20 mM | 0-150 mM | | 磷酸根离子/ Phosphate ion | 0-10 mM | 0-100 mM |
|
【产品特性】
1、本品经利用大肠杆菌 Escherichia coli (E. coli)大规模发酵表达纯化，纯度＞95%，可用于科学研究中降低细胞上清和细胞裂解液的粘度，提高蛋白纯化效率及功能研究，并且可以有效防止细胞治疗和疫苗研究中人外周血单核细胞（PBMC）的结团。
2、 本品以无菌液体酶的形式提供，储存于缓冲液（20 mM Tris-HCl pH8.0，2mM MgCl2，20mM NaCl，50%甘油）中，无色透明液体。
【稳定性】
全能核酸酶在非常广泛的条件下，都能保持很高的稳定性和消化核酸的活性，这使其能够与多种细胞裂解液，如 RIPA，或含有多种离子和非离子型去污剂、还原剂的蛋白提取试剂等兼容。例如：>100mM 的盐酸胍会完全抑制活性，1mM 的 EDTA 会部分抑制活性，5mM 的EDTA 会使活性丧失 90%，1mM 的 PMSF 则不会抑制核酸酶活性。
【使用方法】
1、细胞处理/Cell processing:
a. 贴壁细胞去除培养基，用 PBS 洗后，1mL RIPA 裂解液（或其他哺乳动物细胞裂解液）加1-2 μL全能核酸酶，室温或冰上孵育 5-30 min，收集裂解液，离心取上清即
可进行下游实验。
b. 悬浮细胞离心收集后，在离心管中加 1mL RIPA 裂解液（或其他哺乳动物细胞裂解液）加 1-2 μL 全能核酸酶，室温或冰上孵育 5-30 min，收集裂解液，离心取上清即可进行下游实验。
2、组织样品/Organizational samples： 将 30-100 mg 动物或者植物组织研磨充分后，加入 100-200 μL 裂解液，同时加入加 0.5-1 μL 全能核酸酶，室温或冰上孵育 5-30 min，收集裂解液，离心取上清即可进行下游实验。
3、大肠杆菌或者其他细菌/Escherichia coli or other bacteria： 细菌离心收集后，用裂解液裂解或者研磨破碎后，每 1 mL 加 0.5-1μL 全能核酸酶，室温孵育 30 min，收集裂解液，离心取上清即可进行下游实验。
【用途和作用】
1、 去除疫苗类产品生产过程中的外源性核酸，提高产品的生物安全性；
2、 去除病毒颗粒表面缠绕的核酸，利于病毒颗粒的释放和纯化；
3、 降解细胞裂解液中的核酸，降低液体粘度，提高重组蛋白产量;
4、 处理用于 ELISA、电泳等分析，或柱层析纯化的样品，提高分辨率和回收率。
【注意事项 】
为了您的安全和健康，请穿实验服并戴一次性手套操作。
UltraNuclease, also known as non restrictive endonuclease or broad-spectrum nuclease; It is a non-specific endonuclease derived from Serratia Marcesen that can cleave between any nucleotide in the chain, completely digesting the nucleic acid into 2-5 base length 5 '- monophosphate oligonucleotides. It can degrade various forms of DNA and RNA under a wide range of conditions and is widely used to remove nucleic acids from biological products.
【Properties】
1、This product has been extensively fermented, expressed, and purified using Escherichia coli (E. coli), with a purity of over 95%. It can be used in scientific research to reduce the viscosity of cell supernatants and lysis liquids, improve protein purification efficiency and functional research, and effectively prevent the aggregation of human peripheral blood mononuclear cell (PBMC) in cell therapy and
vaccine research.
2、This product is provided in the form of sterile liquid enzyme and stored in buffer solution (20 mM Tris HCl pH 8.0, 2mM MgCl2, 20mM NaCl, 50% glycerol), a colorless and transparent liquid.
【Stability】
UltraNuclease can maintain high stability and digestive activity of nucleic acids under a wide range of conditions, making them compatible with various cell lysis liquids, such as RIPA, or protein extraction reagents containing multiple ionic and non-ionic detergents and reducing agents. For example, guanidine hydrochloride with a concentration of>100mM will completely inhibit activity, 1mM EDTA will partially inhibit activity, 5mM EDTA will cause a 90% loss of activity, and 1mM PMSF will not inhibit nuclease activity.
【Usage】
1、Cell processing:
a. Remove the culture medium from adherent cells, wash with PBS, and add 1-2 μL UltraNuclease to 1mL of RIPA lysis solution (or other mammalian cell lysis solution) at room temperature or ice for 5-30 minutes, collect the lysis liquid, centrifuge and obtain the supernatant for downstream experiments.
b.After collecting suspended cells by centrifugation, add 1mL of RIPA lysis fluid (or other mammalian cell lysis fluid) and 1-2 μL UltraNuclease to the centrifuge tube at room temperature or ice for 5-30 minutes, collect the lysate, centrifuge, and obtain the supernatant for downstream experiments.
2、Organizational samples： After grinding 30-100 mg of animal or plant tissue thoroughly, add 100-200 μL lysis fluid, while adding 0.5-1 μL UltraNuclease at room temperature or ice for 5-30 minutes, collect the lysis liquid, centrifuge and obtain the supernatant for downstream experiments.
3、Escherichia coli or other bacteria：After bacterial centrifugation collection, they are lysed or ground to pieces with lysis liquid, and 0.5-1μL UltraNuclease is added to every 1 mL, incubate at room temperature for 30 minutes, collect the lysis liquid, centrifuge and take the supernatant for downstream experiments.
【Application】
1、 Remove exogenous nucleic acids from the production process of vaccine products to improve their biological safety;
2、Removing the nucleic acid entangled on the surface of virus particles is beneficial for the release and purification of virus particles;
3、 Degradation of nucleic acids in cell lysis fluid, reduction of liquid viscosity, and increase of recombinant protein production;
4、Process samples used for ELISA, electrophoresis, or column chromatography purification to improve resolution and recovery.
【Attention】
For your safety and health, please wear Lab coat and disposable gloves.
ZJ26F0737177
危险品化学品经营许可证（带存储）