Recombinant Human TRACP/PAP/ACP5 Protein, CF

Catalog # Availability Size / Price Qty
3948-AP-010
Product Details
Citations (1)
FAQs
Reviews

Recombinant Human TRACP/PAP/ACP5 Protein, CF Summary

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.
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.
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
Analysis
Ala22
Predicted Molecular Mass
34 kDa
SDS-PAGE
36-40 kDa, reducing conditions

Product Datasheets

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.

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.

Assay Procedure

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
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.

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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
Entrez Gene IDs
54 (Human); 11433 (Mouse); 25732 (Rat)
Alternate Names
ACP5; TRACP; TRACP-5b; TTRRAP

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Osteopontin protects against lung injury caused by extracellular histones
    Authors: G Kasetty, P Papareddy, RKV Bhongir, MN Ali, M Mori, M Wygrecka, JS Erjefält, A Hultgårdh-, L Palmberg, H Herwald, A Egesten
    Mucosal Immunol, 2019;12(1):39-50.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Dephosphorylation

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