Human Neutrophil Elastase/ELA2 Antibody

(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human ELA2 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 950317
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant human ELA2
    Met1-Asn252
    Accession # P08246
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    3-25 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
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.
Data Examples
Detection of Human Neutrophil Elastase/ELA2 by Western Blot. 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.
Immunocytochemistry
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 # 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.
Intracellular Staining 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 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.
Immunohistochemistry
Neutrophil Elastase/ELA2 in Human Lymphoma. 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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:
    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.
  • Entrez Gene IDs:
    1991 (Human); 50701 (Mouse)
  • Alternate Names:
    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
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 application: Western Blot.

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Western Blot Human Neutrophil Elastase/ELA2 Antibody MAB91671
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 WB Human Anonymous 06/19/2017
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Western Blot Human Neutrophil Elastase/ELA2 Antibody MAB91671
Western Blot: Human Neutrophil Elastase/ELA2 Antibody [MAB91671]

Summary

ApplicationWestern Blot
Sample TestedPeripheral blood neutrophils
SpeciesHuman

Other Experimental Details

Other Experimental Detailssamples: Human neutrophil lysateBlock: 5% skim milk in PBS-T for 1hr at RTDilution: 1/2000

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