Overview

  • Product name
    Anti-Neutrophil Elastase antibody
    See all Neutrophil Elastase primary antibodies
  • Description
    Rabbit polyclonal to Neutrophil Elastase
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, ICC, IHC-P, WB, IP, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Full length protein. Human leucocyte elastase.

  • Positive control
    • Bone marrow.

Applications

Our Abpromise guarantee covers the use of ab21595 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 20818377
ICC 1/50 - 1/100.
IHC-P 1/20 - 1/200. PubMed: 20837529We recommend antigen retrieval in a steamer or with 0.04% pepsin for 10 minutes at room temperature. See Abreview.
WB 1/200 - 1/500. Predicted molecular weight: 29 kDa.
IP 1/100 - 1/200.
ELISA 1/1000 - 1/2000.

Target

  • Function
    Modifies the functions of natural killer cells, monocytes and granulocytes. Inhibits C5a-dependent neutrophil enzyme release and chemotaxis.
  • Tissue specificity
    Bone marrow cells.
  • Involvement in disease
    Defects in ELANE are a cause of cyclic haematopoiesis (CH) [MIM:162800]; also known as cyclic neutropenia. CH is an autosomal dominant disease in which blood-cell production from the bone marrow oscillates with 21-day periodicity. Circulating neutrophils vary between almost normal numbers and zero. During intervals of neutropenia, affected individuals are at risk for opportunistic infection. Monocytes, platelets, lymphocytes and reticulocytes also cycle with the same frequency.
    Defects in ELANE are the cause of neutropenia severe congenital autosomal dominant type 1 (SCN1) [MIM:202700]. SCN1 is a disorder of hematopoiesis characterized by a maturation arrest of granulopoiesis at the level of promyelocytes with peripheral blood absolute neutrophil counts below 0.5 x 10(9)/l and early onset of severe bacterial infections.
  • Sequence similarities
    Belongs to the peptidase S1 family. Elastase subfamily.
    Contains 1 peptidase S1 domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • Bone marrow serine protease antibody
    • ELA2 antibody
    • ELANE antibody
    • Elastase 2 antibody
    • Elastase 2 neutrophil antibody
    • Elastase neutrophil expressed antibody
    • Elastase-2 antibody
    • ELNE_HUMAN antibody
    • GE antibody
    • Granulocyte derived elastase antibody
    • HLE antibody
    • HNE antibody
    • Human leukocyte elastase antibody
    • Leukocyte elastase antibody
    • Medullasin antibody
    • NE antibody
    • Neutrophil elastase antibody
    • PMN E antibody
    • PMN elastase antibody
    • Polymorphonuclear elastase antibody
    • SCN1 antibody
    see all

Images

  • ab21595 staining Neutrophil Elastase in rat cornea tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    The section was deparaffinized with ethanol and the antigen was retrieved using a steamer. ab21595 was used at a 1/200 dilution, the secondary was an anti-rabbit IgG 1/5,000 also from Abcam. The DAB staining was performed using a DAB Peroxidase Substrate kit. The slides were counterstained with hematoxylin.
  • ab21595 staining neutrophil elastase (NE, green), DNA stained with propidium iodide (PI, red) in COPD sputum by ICC/IF (Immunocytochemistry/immunofluorescence). The sputum matrix contains condensed matrix traversed by thin neutrophil extracellular traps (NETs) strands associated with bacteria (arrowheads), non-activated polymorphonulear neutrophils (PMNs) (open arrow) and NETotic PMNs (solid arrow); asterisks indicate fully spread NETs.Cells were blocked with 10% normal goat seum in PBS containing 10 mM glycine and 0.2% Tween 20. Samples were incubated with primary antibody (1/50). ab96883 (1/100) was used as the secondary antibody.

  • ab21595 at 1/20 dilution staining rat liver tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The rat was challenged with LPS at 100ug/Kg i.p. The tissue sections were formaldehyde fixed and incubated with the antibody for 2 hours. A biotinylated goat anti-rabbit IgG antibody was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Herrmann JB  et al. Complement C5a Receptor 1 Exacerbates the Pathophysiology ofN. meningitidisSepsis and Is a Potential Target for Disease Treatment. MBio 9:N/A (2018). IHC-P . Read more (PubMed: 29362231) »
  • Malcolm KC  et al. Neutrophil killing of Mycobacterium abscessus by intra- and extracellular mechanisms. PLoS One 13:e0196120 (2018). Read more (PubMed: 29672589) »
See all 48 Publications for this product

Customer reviews and Q&As

1-10 of 25 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human ovarian cystedenocarcinoma tissue sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: bicarbonate buffer pH 6.0
Permeabilization
No
Specification
human ovarian cystedenocarcinoma tissue sections
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 03 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Mouse Tissue sections (skin)
Specification
skin
Permeabilization
Yes - tritonX-100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 08 2014

Answer

According to the datasheet, this antibody is a non-purified whole serum product. That means, along with Ascites and cell culture supernatant products, the antibody concentration cannot be determined as there are too many other proteins in this solution as well.
However, we have a table under our Technical FAQs (What concentration of primary antibody should I use?) which explains how much specific antibody can be expected in the non-purified products.
In addition, it will give you a starting point for the dilution of your antibody in different applications.

Read More

Answer

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab21595. I would also appreciate if you can confirm some further details: Sometimes the antibody may react with your blocking solution to create this type of issue. If you have not yet changed this, we would suggest the use of an alternative blocking solution such 3-5% BSA in place of milk. Have you increased the stringency of your washes by increasing time and frequency as well as the addition of a detergent such a Tween? Does the blot glow when adding the ECL reagents? If so we recommend letting the blot rest for some time (up to 1 hour) before exposure. Should the suggestions not improve the results, please do let me know.

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Answer

The application IHC-Fr in mouse was added for two different reasons for these antibodies. ab21595 was validated for use in mouse in IHC-Fr following an abreview and the images were visualised using a secondary antibody conjugated to Alexa Fluor 594. ab68672 was validated for use in mouse in IHC-Fr following a publication:

Elliott CG et al. Periostin modulates myofibroblast differentiation during full-thickness cutaneous wound repair. J Cell Sci 125:121-32 (2012), Supplementary Material, figure S1. Detection was with peroxidase-conjugated secondary antibodies.

Either one of these antibodies would be suitable and would be guaranteed to function in IHC-Fr in mouse.

Read More

Answer

Vielen Dank für Ihre Email.

Frau Zeeck ist in Ihren wohlverdienten Urlaub und deshalb muss ich Ihnen heute die schlechte Nachricht überbringen, dass wir keine anderen Antikörper haben, die bovine Neutrophile Elastase erkennen.

Leider muss ich auch bestätigen, dass auch unsere Antikörper gegen den Standard- Marker für Neutrophile Ly6G nicht für Rind geeignet sind.

Bitte lassen Sie mich wissen, ob Sie noch einen anderen Marker für praktikabel halten und ich schaue gerne nach, ob wir einen Antikörper haben, der möglicher weise auch Rind erkennt.

Wir freuen uns auf den Review und möchten Ihnen versichern, dass wir auch negative Reviews veröffentlichen und für hilfreich und informative ansehen.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse lung)
Specification
Mouse lung
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Nov 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Skin)
Specification
Skin
Fixative
Formaldehyde
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Prot K
Permeabilization
No
Blocking step
BGB as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 31 2012

Answer

Thank you for your enquiry regarding ab21595.

This antibody is not affinity purified but whole antiserum so the concentration if IgG is not determined. However, it is normally between a range of 10-15 mg/ml.

As the datasheet suggests we normally provide the recommended dilution factor(s) for each tested applications:

ICC: 1/50 - 1/100,

IHC-P: 1/20 - 1/200,

WB: 1/200 - 1/500,

IP: 1/100 - 1/200,

ELISA: 1/1000 - 1/2000

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

1-10 of 25 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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