Recombinant Human Prostatic Acid Phosphatase/ACPP, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Prostatic Acid Phosphatase/ACPP, CF Summary

• Details of Functionality: Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >100,000 pmol/min/ug, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Prostatic Acid Phosphatase/ACPP protein
  Met1-Gln379, with a C-terminal 6-His tag
• Accession #: P15309
• N-terminal Sequence: Lys33
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ACPP
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 41 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: 43-55 kDa, reducing conditions

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 under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Assay Procedure:
  • Assay Buffer: 50 mM NaOAc, pH 4.5
  • Recombinant Human Prostatic Acid Phosphatase/ACPP (rhACPP) (Catalog # 6240-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 rhACPP 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 rhACPP 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 5 minutes.
  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:
  • rhACPP: 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 Prostatic Acid Phosphatase/ACPP, CF

• acid phosphatase, prostate
• ACP3
• ACP-3
• ACPP
• EC 3.1.3.2
• PAP
• Prostatic Acid Phosphatase
• prostatic acid phosphotase

Background

Prostatic Acid Phosphatase (ACPP) catalyzes the hydrolysis of a variety of phosphate monoesters, including phosphorylated proteins (1). The activity optimum of ACPP is in the pH range of 4 - 6, and the activity is inhibited by L(+)-tartrate. ACPP expression levels are highest in the prostate, with much lower expression in most other tissues. ACPP is a type I integral membrane protein of the plasma membrane and lysosomes, and a secreted form also exists (2). The concentration of ACPP is elevated in the circulation of prostate cancer patients, making the enzyme a marker for the progression of prostate cancer (3). Cellular ACPP has been shown to be a protein tyrosine phosphatase (4). Protein substrates include the epidermal growth factor receptor and HER-2 (5). Cellular ACPP is considered to function as a tumor suppressor (5).

1. Apostol, I. et al. (1985) Acta. Biochim. Pol. 32:187.
2. Schroeder, B. et al. (2007) Traffic 8:1676.
3. Gutman, E.B. et al. (1936) Am. J. Cancer 28:485.
4. Lin, M.F. and G.M. Clinton (1986) Biochem. J. 235:351.
5. Veeramani, S. et al. (2005) Endocr. Relat. Cancer 12:805.

Bioinformatics

• Gene Symbol: ACPP
• Uniprot:
  • P15309(Human)