Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF

R&D Systems | Catalog # 11143-SE

His-tag
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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse u-Plasminogen Activator (uPA) Protein (11143-SE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

NP_032899.1

Applications

Enzyme Activity
View all u-Plasminogen Activator (uPA)/Urokinase Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse u-Plasminogen Activator (uPA)/Urokinase protein
Gly21-Phe433, with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Lys157, Ile180, Met361

Predicted Molecular Mass

18 kDa (long A chain), 3 kDa (short A chain), 29 kDa (B chain)

SDS-PAGE

24-33 kDa (short form), under non-reducing conditions

Activity

Measured by its ability to cleave a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC).
The specific activity is >1700 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF

Recombinant Mouse u‑Plasminogen Activator (uPA)/Urokinase His-tag Protein Enzyme Activity.

Recombinant Mouse u-Plasminogen Activator (uPA)/Urokinase His-tag (Catalog # 11143-SE) is measured by its ability to cleave a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC).

Formulation, Preparation, and Storage

11143-SE
Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl and CaCl2.
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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: u-Plasminogen Activator (uPA)/Urokinase

Urokinase Plasminogen Activator (uPA), also known as u-plasminogen activator or urokinase, is a highly-specific serine protease from the peptidase S1 family that cleaves plasminogen to form plasmin making it a key player in the plasminogen activator (PA) system (1, 2). In cancer, the PA system plays a commanding role in tumor growth, angiogenesis, tumor cell invasion, migration, and metastasis. Expression of uPA is minimal in normal cells but is increased several fold in tumor cells by extracellular stimuli elevated in cancer (3) and corresponds to poor outcomes in several types of cancer (2, 4-7). Therefore, uPA has been identified as an excellent target for therapeutic development through inhibition of protease activity or though inhibition of uPA-dependent signaling while in complex with uPA receptor (uPAR) (2, 7). The pro-enzyme of uPA is synthesized with an N-terminal signal peptide and processed into an active disulfide-linked two-chain molecule (2, 7-10). For mouse uPA, the B chain starting at Ile180 corresponds to the catalytic domain. Two forms of the A chain exist, one starting at Gly21 (the long form) and the other at Lys157 (the short form). While the B chain is common for both forms, the long and short A chains are unique to expected 47 kDa and 32 kDa two-chain forms, respectively. The long A chain contains an EGF-like domain and the kringle domain. The long A domain is reported as responsible for the binding of the uPA receptor (uPAR) (2, 7).

References

  1. Ellis, V. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. eds., Academic Press, San Diego, pp.1677. 
  2. Mahmood, N. et al. (2018) Front Oncol. 8:24.
  3. Nagamine, Y. et al. (2005) Thromb. Haemost. 93:661.
  4. Duffy, M. and C. Duggan. (2004) Clin. Biochem. 37:541.
  5. Pappot, H. et al. (2006) Lung Cancer 51:193.
  6. Taubert, H. et al. (2010) Br. J. Cancer 102:731.
  7. Masucci, M.T. et al. (2022) Cancers. 14:498.
  8. Riccio, A. et al. (1985) Nucleic Acids Res. 13:2759.
  9. Nagai, M. et al. (1985) Gene 36:183.
  10. Jacobs, P. et al. (1985) DNA 4:139.

Long Name

Urokinase-type Plasminogen Activator

Alternate Names

PLAU, uPA, uPlasminogen Activator, Urokinase

Entrez Gene IDs

5328 (Human); 18792 (Mouse); 102135886 (Cynomolgus Monkey)

Gene Symbol

PLAU

UniProt

NP_032899.1

Additional u-Plasminogen Activator (uPA)/Urokinase Products

Product Documents for Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF

For research use only

Citations for Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF

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Protocols

View specific protocols for Recombinant Mouse u-Plasminogen Activator (uPA) Protein, CF (11143-SE):

Materials
  • Assay Buffer: 50 mM Tris, 0.01% (v/v) Tween® 20, pH 8.5
  • Recombinant Mouse u-Plasminogen Activator (uPA)/Urokinase (rmuPA) (Catalog # 11143-SE)
  • Substrate: Z-Gly-Gly-Arg-AMC (Bachem, Catalog # I-1140), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax M5 by Molecular Devices) or equivalent
  1. Dilute rmuPA to 1 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 μL of 1 ng/µL rmuPA into a black well plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL of Assay buffer and 50 µL of 2 mM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

    

*Adjusted for Substrate Blank
 **Derived from calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A9891)
Per Well:
  • rmuPA: 0.05 µg
  • Substrate: 1 mM

























FAQs

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Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail