Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Intracellular Staining by Flow Cytometry

Label

Alexa Fluor 488 (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Monoclonal Mouse IgG1 Clone # 950317
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Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human ELA2
Met1-Asn252
Accession # P08246

Specificity

Detects human ELA2 in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody

Detection of Neutrophil Elastase/ELA2 antibody in THP-1 Human Cell Line antibody by Flow Cytometry.

Detection of Neutrophil Elastase/ELA2 in THP‑1 Human Cell Line by Flow Cytometry.

THP-1 human acute monocytic leukemia cell line treated with 3 µM monensin for 3 hours was stained with Mouse Anti-Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Monoclonal Antibody (Catalog # IC91671G, filled histogram) or isotype control antibody (Catalog # IC002G, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Membrane-associated Proteins.

Applications for Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

0.25-1 µg/106 cells
Sample: THP‑1 human acute monocytic leukemia cell line treated with monensin were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Reviewed Applications

Read 2 reviews rated 4.5 using IC91671G in the following applications:

Spectra Viewer

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Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

Spectra Viewer

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Protect from light. Do not freeze.
  • 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).

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

Additional Neutrophil Elastase/ELA2 Products

Product Documents for Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody


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.

For research use only

Citations for Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody

Customer Reviews (2)

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Showing  1 - 2 of 2 reviews Showing All
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  • Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Blood lymphocytes
    Species: Human
    Verified Customer | Posted 09/06/2024
    Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody IC91671G
  • Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody
    Name: Vincent Giacalone
    Application: Flow Cytometry
    Sample Tested: bronchoalveolar lavage fluid and Blood monocytes
    Species: Human
    Verified Customer | Posted 05/15/2020
    I used this antibody to stain purified leukocytes from blood and BAL fluid. Here airway macrophages are shown having scavenged extracellular elastase and are NE+, compared to NE- blood monocytes, demonstrating an approx. 20x median fluorescence intensity increase. It provides reliable fluorescence when staining cells and can be used to create consistent single-stained bead compensation controls, providing that accurate calibration of the flow cytometer is performed prior to sample acquisition. Signal loss following fixation is minimal when stored at 4 degrees for up to 2 weeks, but I have not assessed samples stored longer than that.
    Human Neutrophil Elastase/ELA2 Alexa Fluor® 488-conjugated Antibody IC91671G

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