Recombinant Human PRL-2/PTP4A2 Protein, CF

Catalog # Availability Size / Price Qty
6694-PT-050
Product Details
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Recombinant Human PRL-2/PTP4A2 Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<0.01 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 >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 #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
20 kDa
SDS-PAGE
20-22 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.

6694-PT

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

Assay Procedure

Materials
  • 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
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: PRL-2/PTP4A2

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).

References
  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.
Long Name
Phosphatase of Regenerating Liver 2
Entrez Gene IDs
8073 (Human)
Alternate Names
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

FAQs

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