Human Neutrophil Elastase/ELA2 Antibody (950302)

R&D Systems | Catalog # MAB91672

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA, Immunocytochemistry

Cited:

Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 950302
View all Neutrophil Elastase/ELA2 Primary Antibodies »

Product Specifications

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human Neutrophil Elastase/ELA2 Antibody (950302)

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 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 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 # 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 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 # 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.
Human Neutrophil Elastase/ELA2 Antibody in ELISA Standard Curve.

Human Neutrophil Elastase/ELA2 ELISA Standard Curve.

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).
Detection of jd1tnkhf1wwwy2olcudc.jpg by Immunohistochemistry

Detection of jd1tnkhf1wwwy2olcudc.jpg by Immunohistochemistry

Neutrophils are involved in Aspergillus (A). fumigatus infection. Lung tissues obtained from patients with mold infection in the lungs were stained with (A) hematoxylin and eosin (H&E) staining, (B) immunohistochemical staining for human neutrophil elastase (ELA2), and (C) methenamine silver staining (MST). (D) Immunofluorescence staining for ELA2 and Calcofluor white staining (CWS) in lung tissue samples with Aspergillus spp. infections. Leukocyte percentages (E) and numbers (F) in the peripheral blood of patients with Aspergillus spp. infections. (G) Neutrophil numbers in the lung tissue sections were counted. Ctrl, lung tissue sections free from Aspergillus spp. Infection; P1 and P2, lung tissue sections with Aspergillus spp. infections. HPF, high power field. *p < 0.05. Original magnification × 100. The red arrow indicates neutrophils. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fmicb.2022.837776/full), 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

Neutrophils are involved in Aspergillus (A). fumigatus infection. Lung tissues obtained from patients with mold infection in the lungs were stained with (A) hematoxylin and eosin (H&E) staining, (B) immunohistochemical staining for human neutrophil elastase (ELA2), and (C) methenamine silver staining (MST). (D) Immunofluorescence staining for ELA2 and Calcofluor white staining (CWS) in lung tissue samples with Aspergillus spp. infections. Leukocyte percentages (E) and numbers (F) in the peripheral blood of patients with Aspergillus spp. infections. (G) Neutrophil numbers in the lung tissue sections were counted. Ctrl, lung tissue sections free from Aspergillus spp. Infection; P1 and P2, lung tissue sections with Aspergillus spp. infections. HPF, high power field. *p < 0.05. Original magnification × 100. The red arrow indicates neutrophils. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fmicb.2022.837776/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Neutrophil Elastase/ELA2 Antibody (950302)

Application
Recommended Usage

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human Neutrophil Elastase/ELA2 Monoclonal Antibody (Catalog # MAB91673).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Neutrophil Elastase/ELA2 DuoSet ELISA Kit (Catalog # DY9167-05) for convenient development of a sandwich ELISA.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed THP-1 human acute monocytic leukemia cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymphoma

Western Blot

1 µg/mL
Sample: Human lung tissue and Human bone marrow

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 (950302)

Certificate of Analysis

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Product Specific Notices for Human Neutrophil Elastase/ELA2 Antibody (950302)

For research use only

Citations for Human Neutrophil Elastase/ELA2 Antibody (950302)

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Protocols

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