Recombinant Human TRACP/PAP/ACP5 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human TRACP/PAP/ACP5 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >65,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human TRACP/PAP/ACP5 protein
Ala22-Pro320, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Ala22
Protein/Peptide Type
Recombinant Enzymes
Gene
ACP5
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
34 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
36-40 kDa, reducing conditions
Publications
Read Publication using
3948-AP in the following applications:

Packaging, Storage & Formulations

Storage
Do not freeze.
  • 3 months from date of receipt, 2 to 8 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM NaOAc, pH 5.0
  • Recombinant Human TRACP/PAP/ACP5 (rhACP5) (Catalog # 3948-AP)
  • Substrate: p-Nitrophenyl phosphate (Sigma-Aldrich, Catalog # N2765)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  • NaOH, 0.2 M in deionized water
  1. Dilute rhACP5 to 0.1 μg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. In a plate combine 50 μL of rhACP5 and 50 μL of 2 mM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 2 mM Substrate.
  4. Incubate reaction at room temperature for 10 minutes in the dark.
  5. Add 100 μL of 0.2 M NaOH to stop the reaction and develop the color.
  6. Read (top read) absorbance in endpoint mode at 410 nm.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).

Per Well:
  • rhACP5: 0.005 μg
  • Substrate: 0.5 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TRACP/PAP/ACP5 Protein, CF

  • ACP5
  • HPAP
  • TRACP
  • TRACP5a
  • TRACP5b
  • TRACP-5b
  • TRAP
  • TrATPase
  • TTRRAP

Background

Encoded by the ACP5 gene, Tartrate Resistant Acid Phosphatase (TRACP or TRAP) is also known as Purple Acid Phosphatase (PAP) or Acid Phosphatase 5 (ACP5) (1). The deuced amino acid (aa) sequence of human ACP5 predicts a signal peptide (aa 1 to 21) and a mature chain (aa 22 to 325). R&D Systems’ recombinant human ACP5 consists of aa 22 to 320, without the last 5 residues, RRARP.

ACP5 is expressed at high levels by osteoclasts, macrophages and dendritic cells (2). Two forms, 5a and 5b, circulating in human blood, are derived from different cell types and have different functions. Derived from macrophages and dendritic cells, 5a is a marker of inflammatory conditions. Derived from osteoclasts, 5b is a marker of bone resorption. Compared to 5a, 5b does not contain sialic acid residues, has a higher specific activity and pH optimum, and may be processed into a disulfide-linked dimer (3).

  1. Janckila, A.J. and J.M. Halleen (2003) J. Bone Miner. Res. 18:1892.
  2. Halleen, J.M. et al. (2006) Clin. Lab. 52:499.
  3. Janckila, A.J. et al. (2003) J. Bone Miner. Res. 18:1916.

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Publications for TRACP/PAP/ACP5 (3948-AP)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


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Bioinformatics

Gene Symbol ACP5
Uniprot