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Rabbit Recombinant Monoclonal Neutrophil Elastase antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 1 publication.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (AB219585), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (AB219585), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (AB219585), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (AB219585), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (AB219585), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFFlow Cyt (Intra)IHC-P
Human
Expected
Tested
Tested
Tested

Expected
Expected

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species
Human
Dilution info
-
Notes

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

7 products for Alternative Version

Target data

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

Recommended products

Rabbit Recombinant Monoclonal Neutrophil Elastase antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 1 publication.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR7479
Purification technique
Affinity purification Protein A
Dissociation constant
6 x 10-12 M
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab219585 is the carrier-free version of Anti-Neutrophil Elastase antibody [EPR7479] ab131260.

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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Anti-Neutrophil Elastase antibody [EPR7479] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • Immunocytochemistry/ Immunofluorescence - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Anti-Neutrophil Elastase antibody [EPR7479] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Overlay histogram showing HL-60 cells stained with Anti-Neutrophil Elastase antibody [EPR7479] ab131260 (red line) at 1/20 dilution. The cells were fixed with 80% methanol. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG used under the same conditions. Cells also incubated without primary antibody and secondary antibody (blue line)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Anti-Neutrophil Elastase antibody [EPR7479] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Immunohistochemical analysis of paraffin-embedded Human bone marrow tissue labelling Neutrophil Elastase with unpurified Anti-Neutrophil Elastase antibody [EPR7479] ab131260 at 1/250 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Overlay histogram showing HL60 cells stained with unpurified Anti-Neutrophil Elastase antibody [EPR7479] 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 (Anti-Neutrophil Elastase antibody [EPR7479] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Neutrophil Elastase antibody [EPR7479] ab131260).

  • OI-RD Scanning - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585), expandable thumbnail

    OI-RD Scanning - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    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] - BSA and Azide free (ab219585), expandable thumbnail

    Western blot - Anti-Neutrophil Elastase antibody [EPR7479] - BSA and Azide free (ab219585)

    Neutrophil Elastase Western blot staining using rabbit Anti-Neutrophil Elastase antibody

    This data was developed using Anti-Neutrophil Elastase antibody [EPR7479] ab131260, the same antibody clone in a different buffer formulation.

    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. Anti-Neutrophil Elastase antibody [EPR7479] 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] (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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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