Recombinant Human Active Trypsin 3/PRSS3 Protein, CF
Recombinant Human Active Trypsin 3/PRSS3 Protein, CF Summary
Product Specifications
Ile24-Ser247, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
3714-SE
| Formulation | Supplied as a 0.2 μm filtered solution in HCl, NaCl, CaCl2 and Brij-35. |
| 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.
|
Assay Procedure
- Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
- Recombinant Human Active Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3714-SE)
- 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
- Dilute rhTrypsin 3 to 0.06 µg/mL in Assay Buffer.
- Dilute Substrate to 80 µM in Assay Buffer.
- Load 50 µL of 0.06 µg/mL of rhTrypsin 3 into a plate, and start the reaction by adding 50 µL of 80 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 80 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).
- rhTrypsin 3 (Active form): 0.003 µg
- Substrate: 40 µM
Reconstitution Calculator
Background: Trypsin 3/PRSS3
Human Trypsin 3, encoded by the PRSS3 gene, is also known as mesotrypsin (1). Constituting less than 10% of the total trypsin content in normal pancreatic juice, it is one of the three trypsin isoforms produced by the pancreas (2). Compared to trypsin 1 and 2, one intriguing feature of Trypsin 3 is its resistance to polypeptide trypsin inhibitors, such as the Kunitz-type soybean trypsin inhibitor or the Kazal-type pancreatic secretory trypsin inhibitor. As revealed by the crystal structure, this resistance is likely due to the presence of an arginine residue in place of the highly conserved Gly198 (3). Trypsin 3 is synthesized in the pancreas and secreted into the duodenum lumen, where it is activated by enterokinase. One proposed physiological function of Trypsin 3 is degradation of trypsin inhibitors, which facilitates the digestion of those foods rich in these proteins (4).
- Nyaruhucha, C. N. M. et al. (1997) J. Biol. Chem. 272:10573.
- Rinderknecht, H. et al. (1984) Gastroenterology 86:681.
- Katona, G. et al. (2002) J. Mol. Biol. 315:1209.
- Szmola, R. et al. (2003) J. Biol. Chem. 278:48580.
Citations for Recombinant Human Active Trypsin 3/PRSS3 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 3
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Cystine-knot peptide inhibitors of HTRA1 bind to a cryptic pocket within the active site region
Authors: Li, Y;Wei, Y;Ultsch, M;Li, W;Tang, W;Tombling, B;Gao, X;Dimitrova, Y;Gampe, C;Fuhrmann, J;Zhang, Y;Hannoush, RN;Kirchhofer, D;
Nature communications
Species: N/A
Sample Types: Peptide, Protein, Small Molecule
Applications: Bioassay -
Engineering and Structural Insights of a Novel BBI-like Protease Inhibitor Livisin from the Frog Skin Secretion
Authors: J Yang, C Tong, J Qi, X Liao, X Li, X Zhang, M Zhou, L Wang, C Ma, X Xi, T Chen, Y Gao, D Wu
Toxins, 2022-04-12;14(4):.
Species: Odorrana livida
Sample Types: Peptide
Applications: Bioassay -
Activated hepatic stellate cells are dependent on self-collagen, cleaved by membrane type 1 matrix metalloproteinase for their growth.
Authors: Birukawa N, Murase K, Sato Y, Kosaka A, Yoneda A, Nishita H, Fujita R, Nishimura M, Ninomiya T, Kajiwara K, Miyazaki M, Nakashima Y, Ota S, Murakami Y, Tanaka Y, Minomi K, Tamura Y, Niitsu Y
J Biol Chem, 2014-05-27;289(29):20209-21.
Applications: Enzyme Assay
FAQs
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