Recombinant Human t-Plasminogen Activator/tPA Protein, CF

R&D Systems | Catalog # 7449-SE

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems CHO-derived Recombinant Human t-Plasminogen Activator/tPA Protein (7449-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

CHO

Accession Number

P00750

Applications

Enzyme Activity
View all t-Plasminogen Activator/tPA Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human t-Plasminogen Activator/tPA protein
Met1-Pro562, with a C-terminal 6-His tag
Single-chain proform was expressed, purified, activated with thermolysin to a 2-chain protease and further purified.

Purity

>85%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser36 (Chain A) and Ile311 (Chain B)

Predicted Molecular Mass

31 kDa (Chain A), 29 kDa (Chain B)

SDS-PAGE

30-40 kDa, 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 >190 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 7449-SE in the following applications:

Scientific Data Images for Recombinant Human t-Plasminogen Activator/tPA Protein, CF

Recombinant Human t-Plasminogen Activator/tPA Protein Enzyme Activity

Recombinant Human t-Plasminogen Activator/tPA Protein Enzyme Activity

Recombinant Human t-Plasminogen Activator (Catalog # 7449-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

7449-SE
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and CaCl2.
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.

Background: t-Plasminogen Activator/tPA

PLAT, also known as tissue-type plasminogen activator (tPA), is a secreted serine protease synthesized by endothelial cells (1).  The partially active single chain can be further processed to full activity by plasmin, tissue kallikrein or Factor Xa.  Active PLAT converts plasminogen to plasmin, a fibrinolytic protease, by hydrolyzing an Arg-Val peptide bond in plasminogen.  Unusually high levels of tPA activity can result in excessive bleeding, and low levels of tPA activity can result in thrombosis or embolism.  Human PLAT contains 4 domains; the N-terminal fibronectin type-1 domain, an epidermal growth factor-like domain, two kringle domains and a serine protease catalytic domain.

References

  1. Lijnen H.R. and D. Collen (2004) Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 1684, Academic Press, San Diego.

Long Name

Tissue Plasminogen Activator

Alternate Names

Alteplase, PLAT, Reteplase, T-PA, tPlasminogen Activator

Entrez Gene IDs

5327 (Human); 18791 (Mouse); 25692 (Rat)

Gene Symbol

PLAT

UniProt

P00750

Additional t-Plasminogen Activator/tPA Products

Product Documents for Recombinant Human t-Plasminogen Activator/tPA Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human t-Plasminogen Activator/tPA Protein, CF

For research use only

Citations for Recombinant Human t-Plasminogen Activator/tPA Protein, CF

Customer Reviews for Recombinant Human t-Plasminogen Activator/tPA Protein, CF (1)

5 out of 5
1 Customer Rating
5 Stars
100%
4 Stars
0%
3 Stars
0%
2 Stars
0%
1 Stars
0%

Have you used Recombinant Human t-Plasminogen Activator/tPA Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card
Showing  1 - 1 of 1 review Showing All
Filter By:
Clear Filters X
  • Recombinant Human t-Plasminogen Activator/tPA Protein, CF
    Name: Leslie Priddy
    Application: Cell Adhesion
    Verified Customer | Posted 04/01/2018

There are no reviews that match your criteria.

Showing  1 - 1 of 1 review Showing All

Protocols

View specific protocols for Recombinant Human t-Plasminogen Activator/tPA Protein, CF (7449-SE):

Materials
  • Assay Buffer: 50 mM Tris, 0.01% Tween-20, pH 8.5
  • Recombinant Human t‑Plasminogen Activator/tPA  (rhPLAT) (Catalog # 7449-SE)
  • Fluorogenic Peptide 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 Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhPLAT to 2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Load 50 µL of the 2 ng/µL rhPLAT into a black well plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 200 µM Substrate without any rhPLAT.
  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 using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A9891).

  • rhPLAT: 0.1 µg
  • Substrate: 100 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes