Recombinant Human PRL-1/PTP4A1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human PRL-1/PTP4A1 Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >0.35 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human PRL-1/PTP4A1 protein
Ala2-Gln173, with an N-terminal Met and 7-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
PTP4A1
Purity
>80%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
21 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
24 kDa, reducing conditions

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl, Betamercaptoethanol and Brij-35.
Purity
>80%, 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-1/PTP4A1 (rhPTP4A1) (Catalog # 8490-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 rhPTP4A1 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 rhPTP4A1 and dilute Substrate in microtubes. Include an Enzyme Control by combining dilute rhPTP4A1 with twice the volume of 0.2 M NaOH, mix briefly, then add a volume of dilute Substrate equivalent to the volume of rhPTP4A1.  The Enzyme Control will have 2x the volume of the reaction mixture.
  4. Incubate Reactions and Enzyme 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 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:
  • rhPTP4A1: 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-1/PTP4A1 Protein, CF

  • EC 3.1.3.48
  • HH72
  • PRL1
  • PRL-1
  • PRL-1DKFZp779M0721
  • Protein tyrosine phosphatase IVA1
  • protein tyrosine phosphatase type IVA 1
  • protein tyrosine phosphatase type IVA protein 1
  • protein tyrosine phosphatase type IVA, member 1
  • Protein-tyrosine phosphatase 4a1
  • Protein-tyrosine phosphatase of regenerating liver 1
  • PTP(CAAX1)
  • PTP(CAAXI)
  • PTPCAAX1PRL1PTP(CAAX1)

Background

Phosphatase of regenerating liver 1 (PRL-1), also known as protein-tyrosine phosphatase 4A1 (PTP4A1), is a member of the PRL subgroup of PTPases (1). It was originally identified as an immediate-early gene in regenerating liver and later shown to be a PTPase that modulates cell growth (2, 3). PRL-1 is widely expressed in human tissues, and the rat enzyme is expressed in neurons and oligodendrocytes of the cerebral cortex, hippocampus, and cerebellum (4, 5). Human PRL-1 shares 100% amino acid sequence identity with mouse and rat PRL-1. The crystal structure of PRL-1 shows that the protein exists as a trimer (6, 7). PRL-1 localizes to the endoplasmic reticulum in interphase cells in a farnesylation-dependent manner, and to centrosomes during mitosis in a farnesylation-independent manner (8, 9). PRL-1 has been implicated in cell migration, invasion, and metastasis, suggesting that it may play a role in cancer (10-13). PRL-1 promotes colorectal cancer cell growth, migration, and invasion in vitro and tumor growth in vivo (14). Additionally, elevated PRL-1 expression is correlated with tumor progression in human hepatocellular carcinoma (15).
  1. Bessette, D.C. et al. (2008) Cancer Metastasis Rev. 27:231.
  2. Mohn, K.L. et al. (1991) Mol. Cell. Biol. 11:381.
  3. Diamond, R.H. et al. (1994) Mol. Cell. Biol. 14:3752.
  4. Dumaual, C.M. et al. (2006) J. Histochem. Cytochem. 54:1401.
  5. Takano, S. et al. (1996) Brain Res. Mol. Brain Res. 40:105.
  6. Sun, J.P. et al. (2005) Biochemistry 44:12009.
  7. Jeong, D.G. et al. (2005) J. Mol. Biol. 345:401.
  8. Zeng, Q. et al. (2000) J. Biol. Chem. 275:21444.
  9. Wang, J. et al. (2002) J. Biol. Chem. 277:46659.
  10. Zeng, Q. et al. (2003) Cancer Res. 63:2716.
  11. Achiwa, H. and J.S. Lazo (2007) Cancer Res. 67:643.
  12. Nakashima, M. and J.S. Lazo (2010) J. Pharmacol. Exp. Ther. 334:627.
  13. Jin, S. et al. (2014) Oncotarget 5:3685.
  14. Zhou, C. et al. (2013) PLoS One 8:e63142.
  15. Lu, J.W. et al. (2012) Clin. Transl. Oncol. 14:287.

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Bioinformatics

Gene Symbol PTP4A1
Uniprot