Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF

Catalog # Availability Size / Price Qty
4517-SE-010
Product Details
Citations (3)
FAQs
Supplemental Products
Reviews (1)

Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF Summary

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.
Activity
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.
Source
Mouse myeloma cell line, NS0-derived mouse Neutrophil Elastase/ELA2 protein
Ser27-Arg260, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ser27
Predicted Molecular Mass
27 kDa
SDS-PAGE
34 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

4517-SE

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rmELA2 to 50 µg/mL in Activation Buffer containing 50 µg/mL rmCathepsin C.
  2. Incubate for 2 hours at 37 °C to activate rmELA2.
  3. Dilute active rmELA2 to 1 ng/µL in Assay Buffer.
  4. Dilute Substrate to 200 µM in Assay Buffer.
  5. 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.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Bieth, J.G. (2004) In Handbook of Proteolytic Enzymes. Barrett, A.J., Rawlings, N.D., & Woessner, J.F. eds. pp. 1517.
  2. Owen, C.A. et al. (1995) J. Cell Biol. 131:775.
  3. Wiesner, O. et al. (2005) FEBS letters 579:5305.
Entrez Gene IDs
1991 (Human); 50701 (Mouse)
Alternate Names
Bone marrow serine protease; EC 3.4.21; EC 3.4.21.37; ELA2; ELA2granulocyte-derived elastase; ELANE; elastase 2, neutrophil; elastase, neutrophil expressed; Elastase-2; GE; HLEelastase-2; HNE; Human leukocyte elastase; Leukocyte Elastase; Medullasin; NE; Neutrophil Elastase; PMN elastase; PMN-E; polymorphonuclear elastase; SCN1

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.

3 Citations: Showing 1 - 3
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  1. 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.
    Species: Mouse
    Sample Types: Protein
    Applications: Bioassay
  2. 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.
    Species: Mouse
    Sample Types: Protein
    Applications: Bioassay
  3. Neutrophils mediate insulin resistance in mice fed a high-fat diet through secreted elastase.
    Nat Med, 2012;18(9):1407-12.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

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Reviews for Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF

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Recombinant Mouse Neutrophil Elastase/ELA2 Protein, CF
By Anonymous on 06/24/2016
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