Recombinant Human PPA1 Protein, CF
R&D Systems | Catalog # 6557-PP
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Key Product Details
- R&D Systems E. coli-derived Recombinant Human PPA1 Protein (6557-PP)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
E. coli
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
E. coli-derived human Inorganic Pyrophosphatase/PPA1 protein
Met1-Asn289, with a C-terminal 6-His tag
Met1-Asn289, with a C-terminal 6-His tag
Purity
>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ser2
Predicted Molecular Mass
33 kDa
SDS-PAGE
34-36 kDa
Activity
Measured by its ability to hydrolyze pyrophosphate.
The specific activity is >50,000 pmol/min/μg, as measured under the described conditions.
The specific activity is >50,000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
6557-PP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Inorganic Pyrophosphatase/PPA1
References
- Harold, FM (1966) Bacteriol. Rev. 30:772.
- Nelson, D.L. and Cox, M.M. (2000) Lehninger Principles of Biochemistry, 3rd ed. pp.937. ISBN 1-57259.
- Poole, K.E. and Reeve, J. (2005) Curr. Opin. Pharmacol. 5:612.
- Takahashi, K. et al. (2004) Biochem. Biophys. Res. Commun. 325:203.
- Terkeltaub, R.A. (2001) Am. J. Physiol., Cell Physiol. 281:C1.
- Wu, Z.L. et al. (2010) BMC Biotechnology 10:11.
- Fairchild, T.A. and Patejunas, G. (1999) Biochim. Biophys. Acta 1447:133.
Alternate Names
IOPPP, IPP1, PP1, PPase, SID6-8061
Gene Symbol
PPA1
UniProt
Additional Inorganic Pyrophosphatase/PPA1 Products
Product Documents for Recombinant Human PPA1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human PPA1 Protein, CF
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human PPA1 Protein, CF (6557-PP):
Materials
- Assay Buffer: 25 mM MES, 16 mM MgCl2, pH 7.0
- Recombinant Human Inorganic Pyrophosphatase/PPA1 (rhPPA1) (Catalog # 6557-PP)
- Substrate: Sodium Pyrophosphate (Sigma, Catalog # P8010)
- ]Malachite Green Phosphate Detection Kit (Catalog # DY996)
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute Substrate to 16 mM in deionized water.
- Dilute rhPPA1 to 0.4 µg/mL in Assay Buffer.
- Combine 25 µL of 16 mM Substrate with 25 µL of 0.4 µg/mL rhPPA1. Also create a substrate blank by combining 25 µL of 16 mM substrate with 25 µL of Assay Buffer.
- Incubate reaction and blank for 10 minutes at room temperature.
- Dilute 1 M Phosphate Standard by adding 10 µL of the 1 M Phosphate Standard to 990 µL of deionized water for a 10 mM stock. Continue by adding 10 µL of the 10 mM Phosphate stock to 990 µL of deionized water for a 100 µM stock.
- Prepare standard curve by performing six one-half serial dilutions of the 100 µM Phosphate stock in deionized water. The standard curve has a range of 0.078 to 5 nmol per well.
- Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of deionized water.
- Dilute the incubated reaction and incubated blank 20-fold by adding 950 μL of deionized water to each tube.
- Load 50 µL of the diluted rhPPA1 reaction and substrate blank into the plate.
- Add 30 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
- Add 100 µL deionized water to all wells.
- Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
- Read plate at 620 nm (absorbance) in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Phosphate released* (nmol) x (1000 pmol/nmol) |
| Incubation time (min) x amount of enzyme (µg) x 2* |
*Derived from the phosphate standard curve using linear or 4-parameter fitting and adjusted for Substrate Blank.
**Note: 2 mol of phosphate are produced for every 1 mol of Pyrophosphate consumed.
Per Well:
- rhPPA1: 0.01 µg in reaction, 0.5 ng after 20-fold dilution
- Substrate: 8 mM in reaction, 0.4 mM after 20-fold dilution