Recombinant Human VHR Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Product Details

Recombinant Human VHR Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Measured by hydrolysis of 250 µM 3-O-methyl fluorescein phosphate (OMFP). The specific activity is >1.0 µmol/min/mg, as measured under the described conditions.
E. coli-derived human VHR protein
Ser2-Pro185, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Predicted Molecular Mass
21 kDa
23 kDa, reducing conditions

Product Datasheets

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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.


Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

  • Assay Buffer: 50 mM HEPES, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM DTT, 0.5 mg/mL BSA, pH 7.5
  • Recombinant Human VHR (rhVHR) (Catalog # 1654-VH)
  • Substrate: 3-O-Methyl Fluorescein Phosphate (OMFP) (Sigma, Catalog # M2629), 2.5 mM stock in DMSO
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhVHR to 5 ng/µL in Assay Buffer.
  2. Dilute substrate OMFP to 500 µM in Assay Buffer. The solution will be light yellow.
  3. Load in a clear plate 50 µL of 5 ng/µL rhVHR, and start the reaction by adding 50 µL of 500 µM Substrate. Include a Substrate Blank containing 50 µL of the Assay Buffer and 50 µL of Substrate.
  4. Read at a wavelength of 477 nm (absorbance) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (μmol/min/mg) =

Adjusted Vmax* (OD/min) x well volume (L) x 106 μmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (mg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 27,200 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhVHR: 0.00025 mg
  • Substrate: 250 µM
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.


Background: VHR

Vaccinia Virus VH1-related Phosphatase (VHR), also known as Dual-Specificity Phosphatase 3 (DUSP3), removes inorganic phosphate groups covalently attached to tyrosine, serine and threonine residues in proteins (1). It belongs to a family of phosphatases that selectively dephosphorylate MAP kinases. VHR has been shown to dephosphorylate cellular ERK1, ERK2 and JNK, but not p38 (2, 3). Phosphorylation of VHR at tyrosine 138 by ZAP-70 enhances inhibition of ERK2, suggesting a role for VHR in modulating T-cell receptor responses (3). The enzymatic mechanism for VHR has been studied in detail (4) and its X-ray crystal structure has been characterized (5). Its well-defined biochemistry has made it useful in screening assays for compounds that inhibit phosphatases (6).

  1. Ishibashi, T. et al. (1992) Proc. Natl. Acad. Sci. USA 89:12170.
  2. Todd, J.L. et al. (1999) J. Biol. Chem. 274:13271.
  3. Alonso, A. et al. (2003) Nat. Immunol. 4:44.
  4. Zhang, Z.-Y. et al. (1995) Biochemistry 34:16088.
  5. Yuvaniyama, J. et al. (1996) Science 272:1328.
  6. Ueda, K. et al. (2002) FEBS Lett. 525:48.
Long Name
Vaccinia Virus VH1-related Phosphatase
Entrez Gene IDs
1845 (Human)
Alternate Names
dual specificity phosphatase 3; dual specificity protein phosphatase 3; Dual specificity protein phosphatase VHR; DUSP3; EC; EC; Vaccinia H1-related phosphatase; vaccinia virus phosphatase VH1-related; VHR; VHRserine/threonine specific protein phosphatase


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