Neutrophil Elastase/ELA2 Antibody (950317) [Unconjugated]

Western blot shows lysates of human bone marrow and human lung tissue. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671) 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 was detected in immersion fixed THP-1 human acute monocytic leukemia cell line using Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671) at 5 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 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91671) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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 # MAB91671, 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.

• Reactivity: Hu
• Applications: WB, IHC, CyTOF-ready, Flow, ICC/IF
• Clone: 950317
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Neutrophil Elastase/ELA2 Antibody (950317) [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 5-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Neutrophil Elastase/ELA2 Antibody (950317) [Unconjugated]

• Bone marrow serine protease
• EC 3.4.21
• EC 3.4.21.37
• ELA2
• ELA2granulocyte-derived elastase
• ELANE
• elastase 2, neutrophil
• elastase, neutrophil expressed
• Elastase-2
• GE
• HLEelastase-2
• HNE
• Human leukocyte elastase
• Leukocyte Elastase
• Medullasin
• NE
• Neutrophil Elastase
• PMN elastase
• PMN-E
• polymorphonuclear elastase
• SCN1

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Neutrophil Elastase/ELA2 Antibody (MAB91671)(9)

Publications using MAB91671

• Barbu, E;Mihaila, AC;Gan, AM;Ciortan, L;Macarie, RD;Tucureanu, MM;Filippi, A;Stoenescu, AI;Petrea, SV;Simionescu, M;Balanescu, SM;Butoi, E; The Elevated Inflammatory Status of Neutrophils Is Related to In-Hospital Complications in Patients with Acute Coronary Syndrome and Has Important Prognosis Value for Diabetic Patients International journal of molecular sciences 2024-05-08 [PMID: 38791147] (Western Blot, Immunocytochemistry, Human)
• Wang J, Blalock S, Levitan G et al. Biological mechanisms of infection resistance in tissue engineered blood vessels compared to synthetic expanded polytetrafluorethylene grafts JVS-Vascular Science 7/14/2023 [PMID: 37662589]
• Jatin Narang, Samreen Jatana, András K. Ponti, Ryan Musich, Joshua Gallop, Angela H. Wei et al. Abnormal thrombosis and neutrophil activation increase hospital-acquired sacral pressure injuries and morbidity in COVID-19 patients Frontiers in Immunology 3/21/2023 [PMID: 37026002]
• GJ Pir, A Parray, R Ayadathil, SV Pananchikk, FA Mir, I Muhammad, A Abubakar, N Amir, S Hussain, KH Haroon, A Muhammad, Y Imam, SN Patro, N Akhtar, A Zakaria, S Kamran Platelet-Neutrophil Association in NETs-Rich Areas in the Retrieved AIS Patient Thrombi International Journal of Molecular Sciences, 2022-11-21;23(22):. 2022-11-21 [PMID: 36430952] (IHC, Human)
• Kostyunin A, Glushkova T, Stasev A et al. Early Postoperative Immunothrombosis of Bioprosthetic Mitral Valve and Left Atrium: A Case Report International journal of molecular sciences 6/16/2022 [PMID: 35743174]
• Frederik Denorme, Irina Portier, John L. Rustad, Mark J. Cody, Claudia V. de Araujo, Chieko Hoki et al. Neutrophil extracellular traps regulate ischemic stroke brain injury Journal of Clinical Investigation 2022-05-16 [PMID: 35358095] (Immunohistochemistry, Human)
• OF McElvaney, T Asakura, SL Meinig, JL Torres-Cas, RS Hagan, C Gabillard, MP Murphy, LB Thorne, A Borczuk, EP Reeves, RE Zumwalt, Y Mikami, TP Carroll, K Okuda, G Hogan, OJ McElvaney, J Clarke, NL McEvoy, PW Mallon, C McCarthy, G Curley, MC Wolfgang, RC Boucher, NG McElvaney Protease-anti-protease compartmentalization in SARS-CoV-2 ARDS: Therapeutic implications EBioMedicine, 2022-02-22;77(0):103894. 2022-02-22 [PMID: 35217407] (IHC, Western Blot, Human)
• L Schindler, LCD Smyth, J Bernhagen, MB Hampton, N Dickerhof Macrophage migration inhibitory factor (MIF) enhances hypochlorous acid production in phagocytic neutrophils Redox Biology, 2021-03-30;41(0):101946. 2021-03-30 [PMID: 33823474] (ICC, Human)
• AJ Jang, JH Lee, M Yotsu-Yama, J Park, S Kye, RL Benza, MJ Passineau, YJ Jeon, T Nyunoya A Novel Compound, "FA-1" Isolated from Prunus mume, Protects Human Bronchial Epithelial Cells and Keratinocytes from Cigarette Smoke Extract-Induced Damage Sci Rep, 2018-07-31;8(1):11504. 2018-07-31 [PMID: 30065307] (Western Blot, Human)

Bioinformatics

• Uniprot:
  • P08246()