Trappin-2 is the human member of the trappin gene family that contains SLPI (1). Trappin-2 consists of an N-terminal transglutaminase substrate domain (residues 23‑60) and a C-terminal four-disulfide core or whey acidic protein (WAP) domain (residues 72‑117). Elafin or ESI (elastase-specific inhibitor) and SKALP (skin‑derived anti‑leucoproteinase) are alternative names for Trappin-2 and reflect its protease targets. However, elafin and SKALP sometimes correspond only to the processed form that contains the C-terminal WAP domain of the molecule, which can be isolated naturally. The recombinant human Trappin-2 corresponds to the full‑length form (residues 23‑117), which migrates as two protein bands under SDS-PAGE due to an unidentified mechanism. In addition to its ability to inhibit human neutrophil elastase, it can also be used as a substrate for transglutaminases.
Recombinant Human Trappin-2/Elafin Protein, CF
R&D Systems | Catalog # 1747-PI
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Trappin-2/Elafin Protein (1747-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
NS0
Accession Number
Applications
Inhibition Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Trappin-2/Elafin protein
Ala23-Gln117, with a C-terminal 10-His tag
Ala23-Gln117, 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
Ala23
Predicted Molecular Mass
11 kDa
SDS-PAGE
13 kDa and 16 kDa, reducing conditions
Activity
Measured by its ability to inhibit neutrophil elastase cleavage of the fluorogenic peptide substrate, MeOSuc-Ala-Ala-Pro-Val-7-amido-4-methylcoumarin (MeOSuc-AAPV-AMC).
The IC50 value is <50 nM, as measured under the described conditions.
The IC50 value is <50 nM, as measured under the described conditions.
Formulation, Preparation, and Storage
1747-PI
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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.
|
Background: Trappin-2/Elafin
References
- Schalkwijk, J. et al. (1999) Biochem. J. 340:569.
Long Name
Elastase Inhibitor, Transglutaminase Substrate, Serine Protease
Alternate Names
Elafin, ESI, PI3, SKALP, Trappin2
Entrez Gene IDs
5266 (Human)
Gene Symbol
PI3
UniProt
Additional Trappin-2/Elafin Products
Product Documents for Recombinant Human Trappin-2/Elafin Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Trappin-2/Elafin Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Trappin-2/Elafin Protein, CF
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Protocols
View specific protocols for Recombinant Human Trappin-2/Elafin Protein, CF (1747-PI):
Materials
- Activation Buffer: 50 mM MES, 50 mM NaCl, pH 5.5
- Assay Buffer: 50 mM Tris, 1.0 M NaCl, 0.05% (w/v) Brij-35, pH 7.5
- Recombinant Human Trappin-2/Elafin (rhTrappin-2) (Catalog # 1747-PI)
- Recombinant Mouse Neutrophil Elastase/ELA2 (rmELA2) (Catalog # 4517-SE)
- Recombinant Mouse Active Cathepsin C/DPPI (rmCathepsin C) (Catalog # 2336-CY)
- Substrate: MeOSuc-Ala-Ala-Pro-Val-AMC (Bachem, Catalog # I-1270), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmELA2 to 50 µg/mL in Activation Buffer containing 50 μg/mL rmCathepsin C.
- Incubate 50 µg/mL rmELA2 at 37 °C for 2 hours to activate.
- Prepare a dilution curve of rhTrappin-2 (MW: 16,000 Da) in Assay Buffer. Make serial dilutions of: 3200, 1600, 800, 600, 400, 200, 100, and 10 nM.
- Dilute activated rmELA2 to 10 µg/mL with Assay Buffer.
- Combine each dilution of rhTrappin-2 with rmELA2 in equal volumes. Include two rmELA2 controls containing equal volumes of rmELA2 and Assay Buffer without any rhTrappin-2.
- Incubate reactions at room temperature for 10 minutes.
- Dilute each reaction five-fold with Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of the diluted incubated curve into a black well plate, and start the reaction by adding 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50% inhibiting concentration (IC50) for rhTrappin-2 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
- The specific activity of rmELA2 at each point may be determined using the following equation (if desired):
|
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:
- rhTrappin-2: 160, 80, 40, 30, 20, 10, 5 and 0.5 nM
- rmELA2: 0.05 µg
- Substrate: 100 µM
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