Recombinant Human PRL-2/PTP4A2 Protein, CF

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

Recombinant Human PRL-2/PTP4A2 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >0.3 pmol/min/μg, as measured under the described conditions.
• Source: E. coli-derived human PRL-2/PTP4A2 protein
  Asn2-Gln167, with an N-terminal Met and 6-His tag
• Accession #: Q12974
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PTP4A2
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 20 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: 20-22 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in HEPES, NaCl and Betamercaptoethanol.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Assay Procedure:
  • Assay Buffer: 50 mM HEPES, 10 mM DTT, pH 7.5
  • Recombinant Human PRL-2/PTP4A2 (rhPRL-2) (Catalog # 6694-PT)
  • Substrate: p-Nitrophenyl phosphate (Sigma, Catalog # N2765), 10 mM stock in deionized water
  • NaOH, 0.2 M in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhPRL-2 to 40 µg/mL in Assay buffer.
  2. Dilute Substrate to 5 mM in Assay buffer.
  3. Prepare reaction mixtures by combining equivalent volumes of dilute rhPRL-2 and dilute Substrate in microtubes. Prepare an Enzyme Control by combining diluted rhPRL-2 with twice the volume of 0.2 M NaOH, mix briefly, then add a volume of Substrate equivalent to volume of rhPRL-2. The Enzyme Control will have 2x the volume of the reaction mixture.
  4. Incubate reactions and controls at 37 °C for 24 hours.
  5. Load 100 µL of Reactions into a plate in triplicate and stop the reactions by adding 100 µL 0.2 M NaOH.
  6. Load 200 µL of Enzyme Controls into a plate in triplicate.
  7. Read plate at 410 nm (absorbance) in endpoint mode.
  8. 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 Enzyme Controls
  **Derived using calibration standard p-Nitrophenol (Sigma, Catalog # 241326)

  Per Well:
  • rhPRL-2: 2 µg
  • pNPP: 1.25 mM

Notes

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

Alternate Names for Recombinant Human PRL-2/PTP4A2 Protein, CF

• EC 3.1.3.48
• HH13
• HH7-2
• HU-PP-1
• HU-PP-1HH13
• OV-1
• OV-1PTP4A
• PRL2
• PRL-2
• PRL-2HH7-2
• PRL2protein tyrosine phosphatase IVA
• protein tyrosine phosphatase IVA2
• protein tyrosine phosphatase type IVA 2
• protein tyrosine phosphatase type IVA, member 2
• Protein-tyrosine phosphatase 4a2
• Protein-tyrosine phosphatase of regenerating liver 2
• PTP(CAAXII)
• PTP4A2
• PTPCAAX2
• PTPCAAX2phosphatase of regenerating liver 2
• ptp-IV1a
• ptp-IV1b

Background

Phosphatase of regenerating liver 2 (PRL-2), also known as protein tyrosine phosphatase 4A2 (PTP4A2) or OV-1, dephosphorylates tyrosine residues. PRL-2, similar to PRL-1 and PRL-3, is an oncogenic protein and considered a candidate cancer biomarker and therapeutic target (1, 2, 3). Over‑expression of PRL-2 correlates with tumor formation and progression, and overall survival depends on estrogen and progestin receptor status (2, 4). In lung cancer cells, PRL-2 promotes cell migration and invasion through an ERK-dependent signaling pathway (5), suppresses p21 promoter activity, and promotes AP-1 activity (6). PRL-2 also is essential for placental development by down-regulating PTEN and activating Akt Protein (7).

1. Stephens, B.J. et al. (2005) Mol. Cancer Ther. 4:1653.
2. Andres, S.A. et al. (2013) Horm. Cancer 4:2083.
3. Rios,P. et al. (2013) FEBS J. 280:505.
4. Hardy, S. et al. (2010) Cancer Res. 70:8959.
5. Wang, Y. and J.S. Lazo. (2012) Oncogene 31:818.
6. Hwang, J.J. et al. (2012) Oncol. Rep. 27:535.
7. Dong, Y. et al. (2012) J. Biol. Chem. 287:32172.

Bioinformatics

• Gene Symbol: PTP4A2
• Uniprot:
  • Q12974(Human)