Human Neutrophil Elastase/ELA2 Antibody (950317)

R&D Systems | Catalog # MAB91671

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 950317
View all Neutrophil Elastase/ELA2 Primary Antibodies »

Product Specifications

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human Neutrophil Elastase/ELA2 Antibody (950317)

Detection of Human Neutrophil Elastase/ELA2 antibody by Western Blot.

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.
Neutrophil Elastase/ELA2 antibody in THP-1 Human Cell Line by Immunocytochemistry (ICC).

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.
Neutrophil Elastase/ELA2 antibody in Human Lymphoma by Immunohistochemistry (IHC-P).

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.
Detection of Neutrophil Elastase/ELA2 antibody in THP-1 Human Cell Line antibody 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.
Detection of Neutrophil Elastase/ELA2 by Western Blot

Detection of Neutrophil Elastase/ELA2 by Western Blot

Form and function of AAT, NE and SLPI in plasma and tracheal aspirates (TA) of SARS-COV-2 ARDS patients.a (i) Western blot analysis of AAT in SARS-COV-2 TAs from individual SARS-CoV-2 ARDS patients labelled P1,P2,P3,P4 and the control group of nsARDS patients labelled P1,P2,P3 demonstrates cleavage of AAT and AAT-NE complexes with significantly more cleavage of AAT seen in the SARS-CoV-2 TA samples. (ii, iii) Western blot analysis of NE in the TAs of the same individuals patients show free NE and cleaved SLPI in SARS-CoV-2 TAs in contrast to the nsARDS control group. b. FRET analysis reveals active NE in all 5 TAs from SARS-COV-2 patients on their 1st day of ICU admission and inhibition with exogenous AAT. Only 1 of the 5 nsARDS TA samples demonstrated active NE which was inhibited by exogenous AAT. Significance was tested for by paired T test; *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35217407), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Neutrophil Elastase/ELA2 by Immunohistochemistry

Detection of Neutrophil Elastase/ELA2 by Immunohistochemistry

The association between excess NETs in aortic tissue and disease severity or prognosis of patients with AAD. (A) Local NETosis was detected in aortic samples from AAD patients, as colocalization of NE with citH3 was observed by confocal microscopy. NE was stained green (Alexa Fluor 488) and citH3 was stained red (Alexa Fluor 647). DAPI staining was used to mark nucleus (blue fluorescence). (B) Compared with normal controls, the number of NETs formed per field in the tissue samples from AAD patients was significantly increased. (C,D) The number of NETs formed per field was positively correlated with the detection risk score of AD and the APACHE II score. (E) The number of NETs formed per field in patients with in-hospital death was significantly higher than that in patients without in-hospital death. (F) The number of NETs formed per field in patients with 1-year survival was significantly lower than that in patients without 1-year survival. **P < 0.01. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fcvm.2021.683445/full), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Neutrophil Elastase/ELA2 by Immunocytochemistry/ Immunofluorescence

Detection of Neutrophil Elastase/ELA2 by Immunocytochemistry/ Immunofluorescence

Abundance of innate immune cells in rHGP.a Anti-CD68 IHC for Desmoplastic and Replacement CRCLM. b Quantitation of the anti-CD68 at multiple regions in CRCLM. c Immunofluorescence showing the colocalization of the Macrophage (CD68) with Arg1 and iNOS. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s44276-023-00007-9), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Neutrophil Elastase/ELA2 Antibody (950317)

Application
Recommended Usage

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
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymphoma

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: THP‑1 human acute monocytic leukemia cell line treated with monensin were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Western Blot

0.1 µg/mL
Sample: Human bone marrow and human lung tissue

Reviewed Applications

Read 2 reviews rated 5 using MAB91671 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

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 (950317)

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Product Specific Notices for Human Neutrophil Elastase/ELA2 Antibody (950317)

For research use only

Citations for Human Neutrophil Elastase/ELA2 Antibody (950317)

Customer Reviews for Human Neutrophil Elastase/ELA2 Antibody (950317) (2)

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  • Human Neutrophil Elastase/ELA2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Periapical granuloma
    Species: Human
    Verified Customer | Posted 11/05/2021
    Human Neutrophil Elastase/ELA2 Antibody (950317) MAB91671
  • Human Neutrophil Elastase/ELA2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Peripheral blood neutrophils
    Species: Human
    Verified Customer | Posted 06/19/2017
    samples: Human neutrophil lysate Block: 5% skim milk in PBS-T for 1hr at RT Dilution: 1/2000
    Human Neutrophil Elastase/ELA2 Antibody (950317) MAB91671

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