Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF
Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF Summary
Ser27-Arg260, with a C-terminal 10-His tag
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 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.
- Activation Buffer: 50 mM MES, 50 mM NaCl, pH 5.5
- Assay Buffer: 50 mM Tris, 1 M NaCl, 0.05% (w/v) Brij-35, pH 7.5
- 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 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).Per Well:
- rmELA2: 0.05 µg
- Substrate: 100 µM
Background: Neutrophil Elastase/ELA2
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.
- Owen, C.A. et al. (1995) J. Cell Biol. 131:775.
- Wiesner, O. et al. (2005) FEBS letters 579:5305.
Citations for Recombinant Mouse Neutrophil Elastase/ELA2 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.
Citations: Showing 1 - 3
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The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell-derived leukocyte elastase.
Authors: Vicuna L, Strochlic D, Latremoliere A, Bali K, Simonetti M, Husainie D, Prokosch S, Riva P, Griffin R, Njoo C, Gehrig S, Mall M, Arnold B, Devor M, Woolf C, Liberles S, Costigan M, Kuner R
Nat Med, 2015;21(5):518-23.
Sample Types: Protein
Regulation of hepatocyte growth factor in mice with pneumonia by peptidases and trans-alveolar flux.
Authors: Raymond W, Xu X, Nimishakavi S, Le C, McDonald D, Caughey G
PLoS ONE, 2015;10(5):e0125797.
Sample Types: Protein
Neutrophils mediate insulin resistance in mice fed a high-fat diet through secreted elastase.
Nat Med, 2012;18(9):1407-12.
Sample Types: Whole Cells
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