The 100μl Reaction
system was prepared: 10μl 10*Reaction Buffer, 1μl 10mM Na4P2O7, 1μl of diluted
Pyrophosphatase, Inorganic or Storage (Dilution) Buffer, as well as
DEPC-treated Water, incubate at 25ºC for 10 minutes. During incubation, 5mM
phosphate standard was diluted with DEPC-treated Water to 0, 2.5, 5, 10, 20,
30, 40, 50μM. Malachite Green Reagent A and Malachite Green Reagent B are mixed
according to the volume ratio of 2:1 (now used), that is, the color developing
agent. Add 200μl phosphate standard or sample with different concentration
(100μl reaction liquid +100μl water) to each hole of the 96-well plate, add
70μl color developer to each hole, gently blow and mix, stand at room
temperature for 30 minutes, and determine the absorbance of 620nm wavelength with
an enzymoleter. According to the concentration of phosphate standard and the
value of A620, the standard curve is drawn, and the phosphate concentration in
the sample is calculated according to the standard curve.
Pyrophosphatase, Inorganic（无机焦磷酸酶）
Pyrophosphatase, Inorganic（无机焦磷酸酶）
Product Details
Product Details
Product Specification
| Species | Yeast |
| Synonyms | Inorganic pyrophosphatase、PPase |
| Concentration | 0.1U/μl |
| Expression System | E.coli |
| Molecular Weight | 33.5 kD |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 20 mM Tris-HCl、100 mM NaCl、1 mM DTT、0.1 mM EDTA 50% Glycerol (pH 8.0 @ 25°C) |
| Stability & Storage |
Store at -25 ~ -15℃
for 2 years
|
| Reference |
[1] Lahti R , Pitk Ranta T , Valve E ,et al. Cloning and characterization of the gene encoding inorganic pyrophosphatase of Escherichia coli K-12.[J].Journal of Bacteriology, 1988, 170(12):5901-7
[2] Satoh T , Watanabe M ,
Nogi S I ,et al. Molecular Cloning, Expression, and Site-Directed
Mutagenesis of Inorganic Pyrophosphatase from Thermus thermophilus HB8[J]. Journal
of Biochemistry, 1998, 124(1):79-88.
|
Background
Pyrophosphatase, Inorganic is a yeast derived
inorganic pyrophosphatase (PPase) expressed by Escherichia coli, which is an
enzyme that catalyzes the conversion of one molecule of pyrophosphate to two
molecules of phosphate ions, causing the inorganic pyrophosphate hydrolysis to
form phosphates. In molecular biology, it can be used to increase the
production of RNA in vitro transcription reactions.
Components
Storage Solution: 0.1U/μl Pyrophosphatase, Inorganic、 20
mM Tris-HCl、100 mM NaCl、1 mM DTT、0.1 mM EDTA 50%
Glycerol (pH 8.0 @ 25°C)
Protocol
Use 1-3 units per ml
in a high yield in vitro RNA synthesis reaction
Unit Definition
Picture
Picture
Bioactivity
SDS-PAGE
1μg (R: reducing condition, N: non-reducing condition).
SEC-HPLC
