Human Neutrophil Elastase/ELA2 Alexa Fluor® 750-conjugated Antibody
Human Neutrophil Elastase/ELA2 Alexa Fluor® 750-conjugated Antibody Summary
Met1-Asn252
Accession # P08246
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
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Preparation and Storage
- 12 months from date of receipt, 2 to 8 °C as supplied.
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).
- Korkmaz, B. et al. (2010) Pharmacol. Rev. 62:726.
- Stein, R.L. et al. (1987) Biochemistry 26:1301.
- Bachovchin, W.W. (1986) Biochemistry 25:7751.
- Garwicz, D. et al. (2005) Haematologica 90:38.
- Owen, C.A. et al. (1995) J. Cell Biol. 131:775.
- Stephan, A. and M. Fabri (2015) Exp. Dermatol. 24:161.
- Carrell, R.W. et al. (1982) Nature 298:329.
- Rice, W.G. and S.J. Weiss (1990) Science 249:178.
- Thompson, R.C. et al. (1986) Proc. Natl. Acad. Sci. USA 83:6692.
- Cooley, J. et al. (2001) Biochemistry 40:15762.
- Wiedow, O. et al. (1990) J. Biol. Chem. 265:14791.
- Taggart, C. et al. (2000) J. Biol. Chem. 275:27258.
- Sinha, S. et al. (1987) Proc. Natl. Acad. Sci. USA 84:2228.
- Okano, K. et al. (1987) J. Biochem. 102:13.
- Makaryan, V. et al. (2015) Curr. Opin. Hematol. 22:3.
Product Datasheets
Product Specific Notices
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
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I used the anti-HNE antibody to stain purified human neutrophils for flow cytometry. After migration towards patient airway fluid, a subset of neutrophils have degranulated and recaptured elastase on the cell surface are now identifiable. I used a red laser with excitation wavelength of 640 nm which is less than optimal for this fluorochrome, but I was was still able to achieve reliable fluorescence signal. Increasing the voltage and/or amount of antibody used would probably yield even better differentiation of the 2 populations. This is a good antibody for assessing surface-bound HNE on myeloid cells as I achieved similar results with macrophages that scavenged extracellular HNE.
