计算溶液所需的质量、体积或浓度。
Pyrophosphatase, Inorganic （yeast）
有货
为什么选择此级别
PharmPure™,医药级 提供 ≥95% 纯度，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
大体系体外转录mRNA 过程中，不可避免的会产生无机焦磷酸盐，这些物质对转录造成了极大的抑制作用，无机焦磷酸酶（ PPase ）可以水解核酸扩增实验中产生的无机焦磷酸盐，促进反应平衡向产物生成端平移，增加产物的量。
PPase（Pyrophosphatase, Inorganic，无机焦磷酸酶）的分子量约为 63kD，可催化无机焦磷酸盐水解生成正磷酸盐：P2O74_+H2O+PPase→2HPO42_。在核酸扩增实验中，PPase 可水解随着反应生成的无机焦磷酸盐，避免其对反应体系的抑制。焦磷酸盐的去除可使反应平衡向产物生成端平移。
本制品为利用大肠杆菌大规模发酵表达的 GMP 级重组无机焦磷酸酶（酵母来源），采用药用规格原辅料生产，并严格控制宿主蛋白质残留、核酸残留等，符合 GMP 规范的产品生产与质量管理规程保障生产过程及所有原辅料可追溯。
质量要求
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遵循以下规范生产
1. ISO 9001:2015, certified facility。
2. 《GMP 附录-细胞治疗产品》国家药品监督管理局。
3. 《人用基因治疗总论-中国药典 2020》国家药典委。
4. USP Chapter <1043>, Ancillary Materials for Cell, Gene, and Tissue-Engineered Products 用于细胞治疗，基因治疗和组织工程产品中的辅料。
5. USP Chapter <92>, Growth Factors and Cytokines Used in Cell Therapy Manufacturing 细胞治疗产品生产过程中细胞因子和生长因子。
6. Ph. Eur. General Chapter 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products 用于生产细胞或基因治疗药物的生物来源原料。
产品特点
1. 水解无机焦磷酸盐 。
2. DNA合成：显著增强 DNA复制能力 。
3. RNA合成：体外转录反应中提高 RNA 产量 。
4. 最佳反应温度为25℃，65℃ 10min可使该酶失活。
产品用途
1. 优化 RNA转录：提高体外转录反应的 RNA产量 。
2. 去除利用焦磷酸盐测定法来进行 SNP基因分型的试剂中的 PPi污染物 。
3. 促进蛋白质、 RNA和 DNA的合成 。
4. 催化 PPi + H2O→2Pi的反应。
5. ssr-pcr optimization：Improve efficiency and increase DNA production.
活性定义
标准反应条件下每分钟催化无机焦磷酸盐生成 1 μmol 磷酸盐所需的酶量定义为 1 个活性单位。
保存体系
20 mM Tris-HCl; 100 mM NaCl; 1 mM DTT; 0.1 mM EDTA; 50% (v/v) Glycerol; pH 8.0。
保存温度
-20±5 ℃。
注意事项
1. 该酶在多种反应 Buffer 中均具有活性，通常该酶在 HDA、LAMP 等实验中，直接加入即可。
2. 该酶的用量在不同的实验中需要优化，通常在 0.05~1U/ml 浓度调整。
3. 该酶的最佳反应温度为 25℃，其在 16~37℃均有活性，65℃ 10min 可使该酶失活。
4. 辅因子：Mg2+对于酶发挥活性是必须的。
Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote the shift of reaction equilibrium to the product generation end, and increase the amount of products.
The molecular weight of PPase (pyrophosphatase, inorganic, inorganic pyrophosphatase) is about 63kd, which can catalyze the hydrolysis of inorganic pyrophosphate to produce orthophosphate: P2O74_+H2O+PPase→2HPO42_. In the nucleic acid amplification experiment, PPase can hydrolyze the inorganic pyrophosphate generated with the reaction to avoid its inhibition on the reaction system. The removal of pyrophosphate can shift the reaction equilibrium to the product generation end.
This product is a GMP level recombinant inorganic pyrophosphatase (yeast source) expressed by large-scale fermentation of E. coli. It is produced with raw and auxiliary materials of medicinal specifications, and the host protein residue and nucleic acid residue are strictly controlled. The product production and quality management procedures in line with GMP specifications ensure that the production process and all raw and auxiliary materials can be traced.
Quality requirements
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Follow the following specifications
1. ISO 9001:2015, certified facility。
2. GMP appendix - cell therapy products State Drug Administration.
3. general introduction to human gene therapy - Chinese Pharmacopoeia 2020, National Pharmacopoeia Committee.
4. USP chapter <1043>, adjuvant materials for cell, gene, and tissue engineered products.
5. USP chapter <92>, growth factors and cytokines used in cell therapy manufacturing.
6. Ph. Eur. General chapter 5.2.12, raw materials of biological origin for the production of cell-based and gene therapy medical products.
Product features
1. hydrolyze inorganic pyrophosphate.
2. DNA synthesis: significantly enhance DNA replication ability.
3. RNA synthesis: increase RNA production in in vitro transcription reaction.
4. The optimal reaction temperature is 25℃, and the enzyme can be inactivated at 65℃ for 10min.
Product usage
1. optimize RNA transcription: improve the RNA yield of in vitro transcription reaction.
2. remove PPI contamination from reagents for SNP genotyping by pyrophosphate assay.
3. promote the synthesis of protein, RNA and DNA.
4. catalyze the reaction of PPI + H2O → 2pi.
5. ssr-pcr optimization：Improve efficiency and increase DNA production.
Activity definition
Catalytic inorganic pyrophosphate formation 1 per minute under standard reaction conditions μ The amount of enzyme required for mol phosphate was defined as 1 active unit.
Preservation system
20 mM Tris-HCl; 100 mM NaCl; 1 mM DTT; 0.1 mM EDTA; 50% (v/v) Glycerol; pH 8.0。
Storage temperature
-20±5 ℃。
Matters needing attention
1. the enzyme has activity in various reaction buffers. Generally, the enzyme can be directly added in HDA, lamp and other experiments.
2. the dosage of the enzyme needs to be optimized in different experiments, usually adjusted at the concentration of 0.05~1u/ml.
3. the optimum reaction temperature of the enzyme was 25 ℃, and it was active at 16~37 ℃, and the enzyme could be inactivated at 65 ℃ for 10min.
4. cofactor: mg2+ is necessary for enzyme activity.
规格
别名
无机焦磷酸酶(酵母)
规格或纯度
医药级, PharmPure™, ≥95%, 0.1U/μl
英文名称
Pyrophosphatase, Inorganic （yeast）
储存条件
-20°C储存
运输条件
超低温运输
纯度
≥95%
名称和识别符
分子类型
小分子
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 B2320504 |
分析证书 | P489053 |
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溶液计算器
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