Recombinant Human PPA1 Protein, CF

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Recombinant Human PPA1 Protein, CF Summary

Product Specifications

>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.
Measured by its ability to hydrolyze pyrophosphate. The specific activity is >50,000 pmol/min/μg, as measured under the described conditions.
E. coli-derived human Inorganic Pyrophosphatase/PPA1 protein
Met1-Asn289, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Predicted Molecular Mass
33 kDa
34-36 kDa

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

  • 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
  1. Dilute Substrate to 16 mM in deionized water.
  2. Dilute rhPPA1 to 0.4 µg/mL in Assay Buffer.
  3. 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.
  4. Incubate reaction and blank for 10 minutes at room temperature.
  5. 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.
  6. 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.
  7. Load 50 µL of each dilution of the standard curve into a plate. Include a curve blank containing 50 μL of deionized water.
  8. Dilute the incubated reaction and incubated blank 20-fold by adding 950 μL of deionized water to each tube.
  9. Load 50 µL of the diluted rhPPA1 reaction and substrate blank into the plate.
  10. Add 30 µL of the Malachite Green Reagent A to all wells. Mix and incubate for 10 minutes at room temperature.
  11. Add 100 µL deionized water to all wells.
  12. Add 30 µL of the Malachite Green Reagent B to all wells. Mix and incubate for 20 minutes at room temperature.
  13. Read plate at 620 nm (absorbance) in endpoint mode.
  14. 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.


Background: Inorganic Pyrophosphatase/PPA1

Inorganic Pyrophosphatases are enzymes that catalyze the hydrolysis of pyrophosphate to two phosphate ions (1). This reaction is highly exergonic, and is utilized in many biochemical pathways, such as DNA synthesis (2) and bone formation (3), to render reactions effectively irreversible (4, 5). Likewise, inorganic pyrophosphatases can be coupled to unfavorable biochemical reactions in order to drive these reactions to completion, such as in the synthesis of activated sulfur donor 3’‑phosphoadenosine-5’-phosphosulfate (PAPS) (6). Two inorganic pyrophosphatases, i.e. PPA1 and PPA2, are found in humans. PPA1 is highly active on pyrophosphate and is ubiquitously expressed (7).

  1. Harold, FM (1966) Bacteriol. Rev. 30:772.
  2. Nelson, D.L. and Cox, M.M. (2000) Lehninger Principles of Biochemistry, 3rd ed. pp.937. ISBN 1-57259.
  3. Poole, K.E. and Reeve, J. (2005) Curr. Opin. Pharmacol. 5:612.
  4. Takahashi, K. et al. (2004) Biochem. Biophys. Res. Commun. 325:203.
  5. Terkeltaub, R.A. (2001) Am. J. Physiol., Cell Physiol. 281:C1.
  6. Wu, Z.L. et al. (2010) BMC Biotechnology 10:11.
  7. Fairchild, T.A. and Patejunas, G. (1999) Biochim. Biophys. Acta 1447:133.
Entrez Gene IDs
5464 (Human); 852296 (Yeast)
Alternate Names
diphosphate phosphohydrolase; EC; inorganic diphosphatase; inorganic pyrophosphatase 1; inorganic pyrophosphatase; IOPPP; IOPPPMGC111556; IPP1; PP1; PPA1; PPase; PPcytosolic inorganic pyrophosphatase; pyrophosphatase (inorganic) 1; pyrophosphatase (inorganic); pyrophosphatase 1; Pyrophosphate phospho-hydrolase; SID6-8061

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.


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