Neutrophil Elastase/ELA2 Antibody (950302) [Unconjugated]

Western blot shows lysates of human lung tissue, human bone marrow, and human kidney tissue (negative control). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91672) 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 # MAB91672) 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 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91672) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC001) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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 (Catalog # MAB91672) 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).

• Reactivity: Hu
• Applications: WB, ELISA, IHC, ICC/IF
• Clone: 950302
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Neutrophil Elastase/ELA2 Antibody (950302) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Neutrophil Elastase/ELA2
  Met1-Asn252
  Accession # P08246
• Specificity: Detects human Neutrophil Elastase/ELA2 in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Western Blot 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.
• 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 (950302) [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.

Bioinformatics

• Uniprot:
  • P08246()