Measured by its ability to cleave the fluorogenic peptide substrate, MeOSuc-Ala-Ala-Pro-Val-7-amido-4-methylcoumarin (MeOSuc-AAPV-AMC). The specific activity is >500 pmol/min/µg, as measured under the described conditions.
Mouse myeloma cell line, NS0-derived mouse Neutrophil Elastase/ELA2 protein Ser27-Arg260, with a C-terminal 10-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
<1.0 EU per 1 μg of the protein by the LAL method.
27 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Recombinant Mouse Active Cathepsin C/DPPI (rmCathepsin C) (Catalog # 2336-CY)
Substrate: MeOSuc-Ala-Ala-Pro-Val-AMC (Bachem, Catalog # I-1270), 10 mM 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 for 2 hours at 37 °C to activate rmELA2.
Dilute active rmELA2 to 1 ng/µL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer.
Load into a plate 50 µL of 1 ng/µL rmELA2, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 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.
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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).
rmELA2: 0.05 µg
Substrate: 100 µM
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF
Bone marrow serine protease
elastase 2, neutrophil
elastase, neutrophil expressed
Human leukocyte elastase
Neutrophil Elastase (ELA2), also known as polymorphonuclear leukocyte elastase, is a serine protease belonging to the chymotrypsin family. Located primarily in the azurophil granules of polymorphonuclear leukocytes, ELA2 function is the degradation of many extracellular matrix macromolecules (1, 2). alpha -1 Antitrypsin (Serpin A1) and secretory leukocyte protease inhibitor (SLPI) have been shown to inhibit ELA2 activity (3). This protein may be involved in lung emphysema, cystic fibrosis, the adult respiratory distress syndrome (ARDS), rheumatoid arthritis, tumor invasion and infectious diseases (1). The purified rmELA2 does not contain the last five amino acid residues of the deduced amino acid sequence (NP_056594).
Bieth, J.G. (2004) In Handbook of Proteolytic Enzymes. Barrett, A.J., Rawlings, N.D., & Woessner, J.F. eds. pp. 1517.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
Review this Product
Be the first to review our Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF and receive a gift card or discount.