Human Neutrophil Elastase/ELA2 Antibody Summary
Accession # P08246
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.
Detection of Human Neutrophil Elastase/ELA2 by Western Blot. Western blot shows lysates of human lung tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for Neutrophil Elastase/ELA2 at approximately 25-30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neutrophil Elastase/ELA2 in THP‑1 Human Cell Line. Neutrophil Elastase/ELA2 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line using Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671R) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Neutrophil Elastase/ELA2 in Human Tonsil. Neutrophil Elastase/ELA2 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671R) at 0.05 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to neutrophils. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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 Monoclonal Antibody (Catalog # MAB91671R, 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.
Preparation and Storage
- 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).
- 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.
No product specific FAQs exist for this product, however you mayView all Antibody FAQs
Cell and Tissue Staining Kits
Reviews for Human Neutrophil Elastase/ELA2 Antibody
There are currently no reviews for this product. Be the first to review Human Neutrophil Elastase/ELA2 Antibody and earn rewards!
Have you used Human Neutrophil Elastase/ELA2 Antibody?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image