Recombinant Human PRL-2/PTP4A2 Protein, CF

Catalog #: 6694-PT Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
6694-PT-050

R&D Systems Recombinant Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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 #

Q12974

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

20 kDa

SDS-PAGE

20-22 kDa, reducing conditions

Product Datasheets

6694-PT

Product Datasheet

COA

SDS

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

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)

Per Well:

rhPRL-2: 2 µg
pNPP: 1.25 mM

Reconstitution Calculator

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

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.

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