Mouse Trypsin-3, encoded by the PRSS3 gene, is also known as mesotrypsin. It consists of a signal peptide (residues 1 to 15), a pro region (residues 16 to 23), and a mature chain (residues 24 to 246) (1). The purified recombinant mouse Trypsin 3 corresponds to the pro form, which can be activated by enterokinase.
Recombinant Mouse Trypsin 3/PRSS3 Protein, CF
R&D Systems | Catalog # 3565-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Trypsin 3/PRSS3 Protein (3565-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
Structure / Form
Pro form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Trypsin 3/PRSS3 protein
Phe16-Asn246, with a C-terminal 10-His tag
Accession # NP_035775
Phe16-Asn246, with a C-terminal 10-His tag
Accession # NP_035775
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
Phe16
Predicted Molecular Mass
26 kDa
SDS-PAGE
29 kDa and 36 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >17,000 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
3565-SE
| Formulation | Supplied as a 0.2 μm filtered solution in HCl and NaCl. |
| 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.
|
Background: Trypsin 3/PRSS3
References
- Halfon, S. et al. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) pp. 1483, Academic Press, San Diego.
Alternate Names
Mesotrypsin, MTG, PRSS3, PRSS4, Trypsin 4, Trypsinogen 15
Gene Symbol
PRSS3
UniProt
Additional Trypsin 3/PRSS3 Products
Product Documents for Recombinant Mouse Trypsin 3/PRSS3 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 Mouse Trypsin 3/PRSS3 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Trypsin 3/PRSS3 Protein, CF (3565-SE):
Materials
- Activation Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% Brij-35 (w/v), pH 8.0
- Recombinant Mouse Trypsin 3/PRSS3 (rmTrypsin 3) (Catalog # 3565-SE)
- Recombinant Human Enteropeptidase/Enterokinase (rhEnterokinase) (Catalog # 1585-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056) 0.6 M in DMSO
- Substrate MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Activate rhEnterokinase with Thermolysin.
- Dilute rhEnterokinase to 100 µg/mL in Activation Buffer.
- Dilute Thermolysin to 3.16 µg/mL in Activation Buffer.
- Mix equal volumes of diluted rhEnterokinase and Thermolysin.
- Incubate at 37 °C for 30 minutes.
- Stop the reaction by adding an equal volume of 20 mM 1,10 Phenanthroline to the reaction tube.
- Activate rmTrypsin 3 with activated rhEnterokinase.
- Dilute activated rhEnterokinase to 2 µg/mL in Assay Buffer.
- Dilute rmTrypsin 3 to 200 µg/mL in Assay Buffer.
- Mix equal volumes of activated rhEnterokinase and rmTrypsin 3.
- Incubate at room temperature for 1 hour.
- Dilute activated rmTrypsin3 to 0.04 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 0.04 µg/mL of rhTrypsin 3 into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
- 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- rmTrypsin 3: 0.002 µg
- Substrate: 10 µM