Name: Anti-Neutrophil Elastase antibody [EPR7479]
SKU: ab131260
Rating: 5 (4 reviews)

Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) is a rabbit monoclonal antibody that is used to detect Neutrophil Elastase in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 30 publications

Related conjugates and formulations

ab219585
Carrier free
Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free
ab305680
Conjugated
PE Anti-Neutrophil Elastase antibody [EPR7479]
ab305681
Conjugated
APC Anti-Neutrophil Elastase antibody [EPR7479]
ab305682
Conjugated
HRP Anti-Neutrophil Elastase antibody [EPR7479]
ab308811
Conjugated
AP Anti-Neutrophil Elastase antibody [EPR7479]
ab310543
Conjugated
Alexa Fluor® 594 Anti-Neutrophil Elastase antibody [EPR7479]
ab310127
Conjugated
Alexa Fluor® 647 Anti-Neutrophil Elastase antibody [EPR7479]
ab312550
Conjugated
Alexa Fluor® 568 Anti-Neutrophil Elastase antibody [EPR7479]
ab312073
Conjugated
Alexa Fluor® 555 Anti-Neutrophil Elastase antibody [EPR7479]
ab317069
Conjugated
Alexa Fluor® 488 Anti-Neutrophil Elastase antibody [EPR7479]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] (AB131260), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/70	Notes For unpurified use at 1/250 - 1/500

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20	Notes For unpurified use at 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000	Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

6 products for Alternative Product

Target data

See full target information ELANE

Function

Serine protease that modifies the functions of natural killer cells, monocytes and granulocytes. Inhibits C5a-dependent neutrophil enzyme release and chemotaxis (PubMed:15140022). Promotes cleavage of GSDMB, thereby inhibiting pyroptosis (PubMed:36899106). Promotes blood coagulation (PubMed:20676107). Through the activation of the platelet fibrinogen receptor integrin alpha-IIb/beta-3, potentiates platelet aggregation induced by a threshold concentration of cathepsin G (CTSG) (PubMed:25211214, PubMed:9111081). Cleaves and thus inactivates tissue factor pathway inhibitor (TFPI) (PubMed:20676107, PubMed:25211214). Capable of killing E.coli but not S.aureus in vitro; digests outer membrane protein A (ompA) in E.coli and K.pneumoniae (PubMed:10947984).

Alternative names

ELA2, ELANE, Neutrophil elastase, Bone marrow serine protease, Elastase-2, Human leukocyte elastase, Medullasin, PMN elastase, HLE

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR7479
Purification technique: Affinity purification Protein A
Dissociation constant: 6 x 10^-12 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Storage information: Stable for 12 months at -20°C

Notes

What is this antibody validated in?

Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Human samples.

What is the molecular weight of Neutrophil Elastase?

Anti-Neutrophil Elastase [EPR7479] (ab131260) specifically detects a band for Neutrophil Elastase (UniProt: P08246) at a molecular weight of 29kDa.

Recommended positive controls

HL60 cell lysate; Human bone marrow tissue

Trusted by the scientific community

Anti-Neutrophil Elastase [EPR7479] (ab131260) was first used in a scientific publication in 2012 and has been cited over 30 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products

We have a range of other formats of antibody clone [EPR7479] also available for your convenience:
ab131260, Carrier free - ab219585, PE - ab305680, APC - ab305681, HRP - ab305682, Alkaline Phosphatase - ab308811, Alexa Fluor® 647 - ab310127, Alexa Fluor® 594 - ab310543, Alexa Fluor® 555 - ab312073, Alexa Fluor® 568 - ab312550, Alexa Fluor® 488 - ab317069

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Neutrophil Elastase also known as neutrophil proteinase 3 has a molecular mass of approximately 30 kDa. It is a serine protease expressed mainly in azurophilic granules of neutrophils. This enzyme breaks down various proteins including elastin at sites of inflammation. Neutrophil Elastase releases during degranulation or through neutrophil extracellular traps (NETs) to combat pathogens.

Biological function summary

The target plays a significant role in neutrophil-mediated tissue remodeling and host defense. It leads to the breaking down of extracellular matrix components contributing to tissue remodeling and repair. Neutrophil Elastase is part of a complex interplay with other proteases like matrix metalloproteinases (MMPs). Its activity is regulated by endogenous inhibitors such as alpha-1 antitrypsin to prevent excessive tissue damage.

Pathways

This protease is important in inflammation and immune response pathways. It plays a role in the complement system by activating C5 a component of the complement cascade. It also acts in conjunction with other proteases such as cathepsin G to modulate immune and inflammatory responses. Neutrophil Elastase's role within these pathways is pivotal for the regulation of inflammation and destruction of pathogens.

Associated diseases and disorders

Individuals with conditions such as chronic obstructive pulmonary disease (COPD) and cystic fibrosis often exhibit dysregulation or excessive activity of Neutrophil Elastase. This excessive activity leads to tissue damage and exacerbates disease progression. The enzyme also interacts with proteins like elastin and collagen in these diseases further contributing to pathological tissue remodeling and damage.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
ab131260 staining Neutrophil Elastase in Human bone marrow tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/5000). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
ab131260 staining Neutrophil Elastase in the HL-60 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/70). Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120(1/500) an Alexa Fluor^®594-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.
Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
Overlay histogram showingHL-60 cells stained with ab131260 (red line) at 1/20 dilution. The cells were fixed with 80% methanol. The secondary antibody used wasa FITC conjugatedgoat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (black line) wasrabbit monoclonal IgG used under the same conditions. Cells also incubated without primary antibody and secondary antibody (blue line)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
ab131260 staining Neutrophil Elastase in Human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/5000). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
Immunohistochemical analysis of paraffin-embedded Human bone marrow tissue labelling Neutrophil Elastase with unpurified ab131260 at 1/250 dilution.
Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
All lanes: Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) at 1/1000 dilution
All lanes: HL-60 Cell Lysate at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 29 kDa
Observed band size: 29 kDa
Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
Overlay histogram showing HL60 cells stained with unpurified ab131260 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab131260, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor^® 488 (IgG H+L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
Secondary antibody - goat anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab6721)
All lanes: Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) at 1/1000 dilution
All lanes: HL60 cell lysate at 10 µg
Secondary
All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 29 kDa
OI-RD Scanning - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260)
Neutrophil Elastase Western blot staining using rabbit Anti-Neutrophil Elastase antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335 is not suitable for detecting human cell lines in western blot assay. ab131260 and Anti-Neutrophil Elastase antibody [RM1077] ab314916 are alternatives for detecting human cell lines.
Neutrophil elastase undergoes proteolytic cleavage to generate fragments (PMID: 25857284).
Lanes 1 - 3: Western blot - Anti-Neutrophil Elastase antibody [EPR28386-66] (Anti-Neutrophil Elastase antibody [EPR28386-66] ab310335) at 1/1000 dilution
Lanes 4 - 6: Western blot - Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) at 1/1000 dilution
Lanes 7 - 9: Western blot - Anti-Neutrophil Elastase antibody [RM1077] (Anti-Neutrophil Elastase antibody [RM1077] ab314916) at 1/1000 dilution
Lanes 1, 4 and 7: HL-60 (human Acute Promyelocytic Leukemia promyeloblast) whole cell lysate at 20 µg
Lanes 2, 5 and 8: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lanes 3, 6 and 9: U937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 30 kDa, 26 kDa
Exposure time: 40s