As a member of the Serpin superfamily of serine protease inhibitors, Serpin E1/PAI‑1 is the principal inhibitor of urokinase‑type plasminogen activator (uPA) and tissue‑type PA (1, 2). As important regulators of extracellular matrix remodeling, uPA and PAI‑1 play a major role in many processes such as angiogenesis, tumor invasion and obesity (2‑4). For example, uPA and PAI-1 are the only tumor prognostic factors validated at the highest level of evidence with regard to their clinical utility in breast cancer (5). The human PAI-1 is initially synthesized as 402 amino acid precursor with a N‑terminal signal peptide (6, 7). PAI‑1 may exist in one of two possible conformations, designated as active or latent (8). The purified rhPAI‑1 is active against rhuPA. The heterogeneity at the N‑terminus of the purified
rhPAI‑1 has been observed before for both the recombinant and native proteins (9).
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Recombinant Human Serpin E1/PAI-1 Protein, CF
R&D Systems | Catalog # 1786-PI
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Key Product Details
- R&D Systems Sf 21 (stably transfected)-derived Recombinant Human Serpin E1/PAI-1 Protein (1786-PI)
- 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
Sf 21 (stably transfected)
Accession Number
Applications
Inhibition Activity
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Product Specifications
Source
Spodoptera frugiperda, Sf 21 (stably transfected)-derived human Serpin E1/PAI-1 protein
Met1-Pro402, with a C-terminal 10-His tag
Met1-Pro402, with a C-terminal 10-His tag
Purity
>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.
N-terminal Sequence Analysis
Ser22 & Val24
Predicted Molecular Mass
44 kDa
SDS-PAGE
43 kDa, reducing conditions
Activity
Measured by its ability to inhibit uPA cleavage of a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC).
The IC50 value is <13 nM, as measured under the described conditions.
The IC50 value is <13 nM, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1786-PI in the following applications:
Scientific Data Images for Recombinant Human Serpin E1/PAI-1 Protein, CF
Recombinant Human Serpin E1/PAI-1 Protein Enzyme Activity
Recombinant Human Serpin E1/PAI-1 (Catalog # 1786-PI) is measured by its ability to inhibit uPA cleavage of a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC).Formulation, Preparation, and Storage
1786-PI
| Formulation | Lyophilized from a 0.2 μm filtered solution in Sodium Acetate, NaCl and CHAPS. |
| Reconstitution | Reconstitute at 500 μg/mL in sterile 50 mM Sodium Acetate and 100 mM NaCl, pH 5.5. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Calculators
Background: Serpin E1/PAI-1
References
- Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
- Stefansson, S. et al. (2003) Curr. Pharm. Des. 9:1545.
- Duffy, M.J. (2002) Clin. Chem. 48:1194.
- Juhan-Vague, I. et al. (2003) J. Thromb. Haemost. 1:1575.
- Harbeck, N. et al. (2002) Clin. Breast Cancer 3:196.
- Pannekoek, H. et al. (1986) EMBO J. 5:2539.
- Ginsburg, D. et al. (1986) J. Clin. Invest. 78:1673.
- Wang, Z. et al. (1996) Biochemistry 35:16443.
- Stromqvist, M. et al. (1994) Protein Expr. Purif. 5:309.
Long Name
Plasminogen Activator Inhibitor
Alternate Names
Nexin, PAI-1, PAI1, PLANH1
Gene Symbol
SERPINE1
UniProt
Additional Serpin E1/PAI-1 Products
Product Documents for Recombinant Human Serpin E1/PAI-1 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.
Product Specific Notices for Recombinant Human Serpin E1/PAI-1 Protein, CF
For research use only
Citations for Recombinant Human Serpin E1/PAI-1 Protein, CF
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Protocols
View specific protocols for Recombinant Human Serpin E1/PAI-1 Protein, CF (1786-PI):
Materials
- Assay Buffer: 50 mM Tris, 0.01% (v/v) Tween® 20, pH 8.5
- Recombinant Human Serpin E1/PAI-1 (rhSerpin E1/PAI-1) (Catalog # 1786-PI)
- Recombinant Human u‑Plasminogen Activator (uPA)/Urokinase (rhuPA) (Catalog # 1310-SE)
- Substrate: Z-Gly-Gly-Arg-AMC (Bachem, Catalog # I-1140)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhuPA to 2 µg/mL in Assay Buffer.
- Prepare a rhSerpin E1/PAI-1 (MW: 44146 Da) dilution curve in Assay Buffer. Make the following serial dilutions: 1500, 750, 375, 187.5, 125, 75, 50, 25, and 12.5 nM.
- Mix equal volumes of the rhSerpin E1/PAI-1 curve and the diluted rhuPA at 2 µg/mL. Include 2 controls containing rhuPA and Assay Buffer only.
- Incubate mixtures at room temperature for 15 minutes.
- After incubation, dilute the mixtures 5 fold in Assay Buffer.
- Dilute the Substrate to 200 µM in Assay Buffer.
- Load 50 µL of the incubated reactions into a plate, and start the reaction by adding 50 µL of 200 µM Substrate to each well.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Determine the 50% inhibiting concentration (IC50) for rhSerpin E1 by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
- The specific activity for rhuPA at each point may be determined using the following formula (if necessary):
|
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 # A-9891).
Per Well:
- rhuPA: 0.010 µg
- rhSerpin E1/PAI-1 curve: 75, 37.5, 18.75, 9.375, 6.25, 3.75, 2.5, 1.25, and 0.625 nM
- Substrate: 100 µM
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Associated Pathways
