Recombinant Human TRACP/PAP/ACP5 Protein, CF

R&D Systems | Catalog # 3948-AP

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human TRACP/PAP/ACP5 Protein (3948-AP)
  • 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

NS0

Accession Number

NP_001602

Applications

Enzyme Activity
View all TRACP/PAP/ACP5 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human TRACP/PAP/ACP5 protein
Ala22-Pro320, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala22

Predicted Molecular Mass

34 kDa

SDS-PAGE

36-40 kDa, reducing conditions

Activity

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.

Formulation, Preparation, and Storage

3948-AP
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: TRACP/PAP/ACP5

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).

References

  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.

Long Name

Tartrate Resistant Acid Phosphatase 5

Alternate Names

ACP5, TRACP-5b, TTRRAP

Entrez Gene IDs

54 (Human); 11433 (Mouse); 25732 (Rat)

Gene Symbol

ACP5

UniProt

NP_001602

Additional TRACP/PAP/ACP5 Products

Product Documents for Recombinant Human TRACP/PAP/ACP5 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human TRACP/PAP/ACP5 Protein, CF

For research use only

Citations for Recombinant Human TRACP/PAP/ACP5 Protein, CF

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Protocols

View specific protocols for Recombinant Human TRACP/PAP/ACP5 Protein, CF (3948-AP):

Materials
  • 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

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