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Recombinant Human Neutrophil Elastase/ELA2 Protein, CF

R&D Systems | Catalog # 9167-SE

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Neutrophil Elastase/ELA2 Protein (9167-SE)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

P08246

Applications

Enzyme Activity
View all Neutrophil Elastase/ELA2 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Neutrophil Elastase/ELA2 protein
Ser28-Asn252, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser28

Predicted Molecular Mass

25 kDa

SDS-PAGE

29-39 kDa, reducing conditions

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 >1,500 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 2 reviews rated 5 using 9167-SE in the following applications:

Formulation, Preparation, and Storage

9167-SE
Formulation Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Neutrophil Elastase/ELA2

Neutrophil Elastase (ELA2, ELANE), also known as HNE, is a chymotrypsin family serine protease that plays a key role in pathogen clearance (1-3). It is expressed by promyelocytes and stored in the intracellular azurophilic granules of polymorphonuclear leukocytes (PMN) (4). These granules fuse with phagosomes, enabling Neutrophil Elastase to participate in the digestion and killing of endocytosed microbes. The enzyme is released by activated neutrophils at sites of inflammation, and it can remain associated with the cell surface or function as a component of neutrophil extracellular nets (NETs) which trap and kill microbial pathogens (5, 6). It also can degrade multiple extracellular matrix proteins including Elastin and Fibronectin (5). In the lung, this activity contributes to pathology in emphysema, cystic fibrosis, and adult respiratory distress syndrome (ARDS) (1). Neutrophil Elastase can be inhibited by Serpin A1/alpha 1-Antitrypsin, SLPI, Serpin B1, and
Trappin-2/Elafin (7-11). Its activity in the lung is increased by exposure to tobacco smoke which inactivates Serpin A1 through methionine oxidation (12). Mature human Neutrophil Elastase shares 73% amino acid sequence identity with mouse and rat Neutrophil Elastase (13, 14). Multiple mutations in the human ELANE gene are causative of severe congenital and cyclic neutropenias (15).

References

  1. Korkmaz, B. et al. (2010) Pharmacol. Rev. 62:726.
  2. Stein, R.L. et al. (1987) Biochemistry 26:1301.
  3. Bachovchin, W.W. (1986) Biochemistry 25:7751.
  4. Garwicz, D. et al. (2005) Haematologica 90:38.
  5. Owen, C.A. et al. (1995) J. Cell Biol. 131:775.
  6. Stephan, A. and M. Fabri (2015) Exp. Dermatol. 24:161.
  7. Carrell, R.W. et al. (1982) Nature 298:329.
  8. Rice, W.G. and S.J. Weiss (1990) Science 249:178.
  9. Thompson, R.C. et al. (1986) Proc. Natl. Acad. Sci. USA 83:6692.
  10. Cooley, J. et al. (2001) Biochemistry 40:15762.
  11. Wiedow, O. et al. (1990) J. Biol. Chem. 265:14791.
  12. Taggart, C. et al. (2000) J. Biol. Chem. 275:27258.
  13. Sinha, S. et al. (1987) Proc. Natl. Acad. Sci. USA 84:2228.
  14. Okano, K. et al. (1987) J. Biochem. 102:13.
  15. Makaryan, V. et al. (2015) Curr. Opin. Hematol. 22:3.

Alternate Names

ELA2, ELANE, Elastase-2, Leukocyte Elastase, Medullasin

Entrez Gene IDs

1991 (Human); 50701 (Mouse)

Gene Symbol

ELANE

UniProt

P08246

Additional Neutrophil Elastase/ELA2 Products

Product Documents for Recombinant Human Neutrophil Elastase/ELA2 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Neutrophil Elastase/ELA2 Protein, CF

For research use only

Citations for Recombinant Human Neutrophil Elastase/ELA2 Protein, CF

Customer Reviews for Recombinant Human Neutrophil Elastase/ELA2 Protein, CF (2)

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    Verified Customer | Posted 08/04/2023
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Protocols

View specific protocols for Recombinant Human Neutrophil Elastase/ELA2 Protein, CF (9167-SE):

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 Human Elastase/ELA2 (rhELA2) (Catalog # 9167-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 stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhELA2 to 50 µg/mL in Activation Buffer containing 50 µg/mL rmCathepsin C.
  2. Incubate for 2 hours at 37 °C to activate rhELA2.
  3. Dilute active rhELA2 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 rhELA2, 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-Methly Coumarin (Sigma, Catalog # A9891).

Per Well:

  • rhELA2: 0.05 µg
  • Substrate: 100 µM

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