Recombinant Human Prostatic Acid Phosphatase/ACPP, CF
Recombinant Human Prostatic Acid Phosphatase/ACPP, CF Summary
Product Specifications
Met1-Gln379, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
6240-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.
|
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
- Dilute rhACPP to 0.1 μg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- 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.
- Incubate reaction at room temperature for 5 minutes.
- Add 100 μL of 0.2 M NaOH to stop the reaction and develop the color.
- Read (top read) absorbance in endpoint mode at 410 nm.
- 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).
- rhACPP: 0.005 μg
- Substrate: 0.5 mM
Reconstitution Calculator
Background: Prostatic Acid Phosphatase/ACPP
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).
- Apostol, I. et al. (1985) Acta. Biochim. Pol. 32:187.
- Schroeder, B. et al. (2007) Traffic 8:1676.
- Gutman, E.B. et al. (1936) Am. J. Cancer 28:485.
- Lin, M.F. and G.M. Clinton (1986) Biochem. J. 235:351.
- Veeramani, S. et al. (2005) Endocr. Relat. Cancer 12:805.
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