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商品編號


Y9380L



品牌


Enzymatics



公司


Enzymatics,inc



公司分類(lèi)


New Products



Size

50 U

商品信息

E. coli
Pyrophosphatase


Product Description

E. coli
pyrophosphatase catalyzes the Mg-dependent reaction of P
2
O
7
-4
+ H
2
O → 2HPO
4
-2
.

Source of Protein
A recombinant
E. coli
strain carrying the
E. coli
Pyrophosphatase gene.

Unit Definition
One unit is the amount of enzyme that will liberate 1 ?mol of phosphate per minute from inorganic pyrophosphate at 37°C and pH 8.5

Molecular weight
19,704 Daltons



Quality Control Analysis

Unit

Activity
The assay is based on that described by Taussky and Shorr (3). Briefly, enzyme dilutions are added to 30mM Tris HCl pH 8.5, 1.5 mM MgCl2 and 1.5mM sodium pyrophosphate. After a 10 min incubation at 37° C, the product formed, 2- orthophosphate, is reacted with ammonium moly
BD
ate to form phosphomoly
BD
ic acid. The phosphomoly
BD
ic acid is then reduced by ferrous sulfate under weak acidic conditions to form a blue color, the absorbance of which is measured at 660nm. The amount of product formed is extrapolated from a phosphate standard curve generated from the ammonium molydate/ferrous sulfate reaction.

Protein

Concentration (OD
280
)
is determined by OD
280
absorbance.

Physical Purity
is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver stain detection. Purity is assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the protein of interest band in the diluted sample.

Single-Stranded

Exonuclease
is determined in a 50 ?L reaction containing a r
ADI
olabeled single-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Exonuclease
is determined in a 50 ?l reaction containing a r
ADI
olabeled double-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Endonuclease
is determined in a 50 ?L reaction containing 0.5 ?g of plasmid DNA and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

E.coli 16S rDNA Contamination
is evaluated using 5 ?L
r
eplicate samples of enzyme solution denatured and screened in a TaqMan qPCR assay for the presence of contaminating
E.coli
genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus.

Supplied in:
20 mM Tris-HCl pH 8.0, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol


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Product Information



E. coli
Pyrophosphatase


Part Number
Y9380L
Price
$213
Concentration
100 U/mL
Unit Size
50 U


SDS

Available on request








Product Specification*


Storage Temperature
-25?C to -15?C


Test

Units Tested

Specification


SDS Purity
n/a
>95%
Specific Activity
n/a
3,500 U/mg
SS Exonuclease
35
<1.0% Released
DS Exonuclease
35
<1.0% Released
DS Endonuclease
35
No Conversion
E.coli
DNA Contamination
35
<10 copies





* For a detailed summary of assay conditions and data, refer to the Quality Controls Analysis section.

References


Cloning and characterization of the gene encoding inorganic pyrophosphatase of Escherichia coli K-12. Lahti R, Pitk?ranta T, Valve E, Ilta I, Kukko-Kalske E, Heinonen J. J Bacteriol. 1988 Dec;170(12):5901-7.

Catalysis by Escherichia coli inorganic pyrophosphatase: pH and Mg2+ dependence. Baykov AA, Hyytia T, Volk SE, Kasho VN, Vener AV, Goldman A, Lahti R, Cooperman BS. Biochemistry. 1996 Apr 16;35(15):4655-61.

A microcolorimetric method for the determination of inorganic phosphorus. Taussky HH, Shorr E. J Biol Chem. 1953 Jun;202(2):675-85




Limitations of Use
This product was developed, manufactured, and sold for in vitro use only. The product is not suitable for admi
NIST
ration to humans or animals. SDS sheets relevant to this product are available upon request.