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A storage reagent for biotinylated inorganic pyrophosphatase

An inorganic pyrophosphatase and biotinylation technology, applied in the biological field, can solve the problem of storage reagents without biotinylated PPase, and achieve the effects of reducing the influence of activity, prolonging storage time, and reducing shedding.

Active Publication Date: 2017-04-05
BEIJING ZHONGKEZIXIN TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many depot or protection reagents for different enzyme proteins have been published or commercially available, currently there is no depot specific for biotinylated PPase

Method used

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  • A storage reagent for biotinylated inorganic pyrophosphatase
  • A storage reagent for biotinylated inorganic pyrophosphatase
  • A storage reagent for biotinylated inorganic pyrophosphatase

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1 A kind of storage reagent of biotinylated inorganic pyrophosphatase

[0025] The following components were included: Tris-HCl 90 mM pH 8.5, Calcium Chloride 75 mM, Magnesium Chloride 75 mM, Ethylene Glycol 35% (v / v) and DTT 4 mM; solvent was deionized water.

Embodiment 2

[0026] Embodiment 2 A kind of storage reagent of biotinylated inorganic pyrophosphatase

[0027] The following components are included: Tris-HCl 85 mM at pH 8.5, calcium chloride 60 mM, magnesium chloride 50 mM, ethylene glycol 30% (v / v) and DTT 3.6 mM; the solvent is deionized water.

Embodiment 3

[0028] Embodiment 3 A kind of storage reagent of biotinylated inorganic pyrophosphatase

[0029] The following components are included: Tris-HCl 80 mM at pH 8.5, calcium chloride 50 mM, magnesium chloride 40 mM, ethylene glycol 28% (v / v) and DTT 3.4 mM; the solvent is deionized water.

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Abstract

The invention provides a storage reagent for biotinylation inorganic pyrophosphatase. The storage reagent is prepared from 60-90 mM of Tris-HCL with the pH being 8.5, 25-75 mM of calcium chloride, 25-75 mM of magnesium chloride, 20-35% ethanediol and 2-4 mM of DTT. The solvent is deionized water. The storage reagent can protect the activity of the biotinylation inorganic pyrophosphatase, the storage time of the biotinylation inorganic pyrophosphatase can be prolonged, the influence caused by repeated freezing thawing on the activity of the biotinylation inorganic pyrophosphatase is lowered, and the phenomenon that biotin labels on the biotinylation inorganic pyrophosphatase fall down can be effectively reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a storage reagent for biotinylated inorganic pyrophosphatase. Background technique [0002] Inorganic pyrophosphatase (Inorganic Pyrophosphatase, PPase, EC3.6.1.1) is a class of enzymes that catalyze the hydrolysis of pyrophosphate (Pyrophosphoric acid, PPi) into orthophosphate. PPase can provide energy for a variety of biosynthetic pathways by decomposing PPi, and push them towards the synthesis direction. For example, glucose and starch synthesis, cellulose synthesis, aminoacyl-tRNA synthesis, fatty acyl-CoA synthesis, DNA and RNA polymerization, etc. PPase exists widely in nature. With the development of technical means, PPase cDNA clones have been successfully obtained from japonicum, rice, barley, tobacco, yeast, Bacillus subtilis, Escherichia coli and thermophilic bacteria. In the catalytic reaction of PPase, it is necessary to combine three or four divalent metal cations, su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/14C12N9/96
CPCC12N9/14C12N9/96C12Y306/01001
Inventor 高静蔡亦梅吴超徐潇张睿王者馥王绪敏殷金龙任鲁风
Owner BEIJING ZHONGKEZIXIN TECH
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