THP-1 human acute monocytic leukemia cell line was stained with Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91673, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). 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 Intracellular Molecules.

Human Neutrophil Elastase/ELA2 ELISA Standard Curve.

Recombinant Human Neutrophil Elastase/ELA2 protein was serially diluted 2-fold and captured by Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91673) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Applications for Human Neutrophil Elastase/ELA2 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

ELISA

This antibody functions as an ELISA capture antibody when paired with Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91672).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Neutrophil Elastase/ELA2 DuoSet ELISA Kit (Catalog # DY9167-05) for convenient development of a sandwich ELISA.

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: THP-1 Human Cell Line

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

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.

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 Human Neutrophil Elastase/ELA2 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Neutrophil Elastase/ELA2 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Neutrophil Elastase/ELA2 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB91673

Key Product Details

Species Reactivity

Validated:

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Applications

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Neutrophil Elastase/ELA2 Antibody

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