Recombinant Human PRL-2/PTP4A2 Protein, CF Summary
Asn2-Gln167, with an N-terminal Met and 6-His tag
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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.
|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.
- 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
- Dilute rhPRL-2 to 40 µg/mL in Assay buffer.
- Dilute Substrate to 5 mM in Assay buffer.
- 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.
- Incubate reactions and controls at 37 °C for 24 hours.
- Load 100 µL of Reactions into a plate in triplicate and stop the reactions by adding 100 µL 0.2 M NaOH.
- Load 200 µL of Enzyme Controls into a plate in triplicate.
- Read plate at 410 nm (absorbance) in endpoint mode.
- 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)
- rhPRL-2: 2 µg
- pNPP: 1.25 mM
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).
- Stephens, B.J. et al. (2005) Mol. Cancer Ther. 4:1653.
- Andres, S.A. et al. (2013) Horm. Cancer 4:2083.
- Rios,P. et al. (2013) FEBS J. 280:505.
- Hardy, S. et al. (2010) Cancer Res. 70:8959.
- Wang, Y. and J.S. Lazo. (2012) Oncogene 31:818.
- Hwang, J.J. et al. (2012) Oncol. Rep. 27:535.
- Dong, Y. et al. (2012) J. Biol. Chem. 287:32172.
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