Recombinant Mouse Active Trypsin 3/PRSS3 Protein, CF Summary
Ile24-Asn246, with an N-terminal Met, 6-His tag and GGSGSGGGSSR linker
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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.
|Formulation||Supplied as a 0.2 μm filtered solution in Citrate, NaCl, CaCl2, Benz, DTT, and Glycerol.|
|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 Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij35, pH 8.0
- Recombinant Mouse Active Trypsin 3/PRSS3 (rmTrypsin 3) (Catalog # 7235-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 rmTrypsin 3 to 0.007 ng/μL in Assay Buffer.
- Dilute Substrate to 20 μM in Assay Buffer.
- Load into a plate 50 μL of 0.007 ng/μL rmTrypsin 3, and start the reaction by adding 50 μL of 20 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 20 μM Substrate.
- Read plate at excitation and emission wavelengths of 320 nm and 405 nm, 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).
- rmTrypsin 3: 0.35 ng (0.00035 μg)
- Substrate: 10 μM
Background: Trypsin 3/PRSS3
Mouse Trypsin 3, encoded by the Prss3 gene, is also known as mesotrypsin and brain trypsin (1). It is one of three trypsin isoforms produced by the pancreas (2). It is secreted into the duodenum lumen, where it is activated by enterokinase. Trypsin 3 differs from Trypsin 1 and 2 because it is resistant to polypeptide trypsin inhibitors such as the Kunitz-type soybean trypsin inhibitor and the Kazal‑type pancreatic secretory trypsin inhibitor (3). It has been proposed that a function of Trypsin 3 is the degradation of trypsin inhibitors to facilitate the digestion of foods rich in these proteins (4). Recombinant mouse Trypsin 3 is expressed and processed from E.coli, which yields the purified mature chain of mouse Trypsin 3.
- Nyarahucha, 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.
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Fluorogenic Peptide Substrates
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