Overview

  • Product name
    Anti-Neutrophil antibody [NIMP-R14]
    See all Neutrophil primary antibodies
  • Description
    Rat monoclonal [NIMP-R14] to Neutrophil
  • Host species
    Rat
  • Tested applications
    Suitable for: ICC, Functional Studies, IHC-P, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to Mouse Neutrophil. Purified BALB/c mouse neutrophils

  • Epitope
    The monoclonal antibody NIMP-R14 is highly specific for murine Ly-6G and Ly-6C.
  • General notes

    The Ly-6G/-6C locus encodes a family of Ly-6 proteins including Ly-6G and Ly-6C. Ly-6 antigens have a molecular weight between 15,000 and 18,000. Ly-6G is together with Ly-6C a component of the myeloid differentiation antigen Gr-1. Ly-6G is a GPI-anchored protein and is a good marker of peripheral neutrophils. Although predominantly presents on neutrophils, it is also expressed on a subset of eosinophils, differentiating premonocytes and plasmacytoid dendritic cells. Ly-6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon gamma, and may play a role in the development and maturation of lymphocytes. It is expressed on bone marrow cells, monocytes/macrophages, neutrophils, endothelial cells, and T cell subsets. Expression of Gr-1 in bone marrow correlates with granulocyte differentiation and maturation. However, the physiological role of Ly-6G alone remains still unclear. The monoclonal antibody NIMP-R14 has been successfully used to stain polymorphonuclear (PMN) cells and monocytes for fluorescent activated cell sorting and in frozen and paraffin sections. Treatment with antibodies in vivo leads to neutropenia and has inhibitory effect on local immune responses. Furthermore, it has been shown to be useful for depletion of neutrophils in mice. It depletes neutrophils as soon as 6 hours after injection and up to 6 days.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituent: 0.1% BSA
  • Concentration information loading...
  • Purity
    Protein G purified
  • Primary antibody notes
    The Ly-6G/-6C locus encodes a family of Ly-6 proteins including Ly-6G and Ly-6C. Ly-6 antigens have a molecular weight between 15,000 and 18,000. Ly6G is together with Ly6c a component of the myeloid differentiation antigen Gr-1. Ly6G a GPI-anchored protein and is a good marker of peripheral neutrophils. Although predominantly presents on neutrophils, it is also expressed on a subset of eosinophils, differentiating premonocytes and plasmacytoid dendritic cells. Ly6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon gamma, and may play a role in the development and maturation of lymphocytes. It is expressed on bone marrow cells, monocytes/macrophages, neutrophils, endothelial cells, and T cell subsets. Expression of Gr-1 in bone marrow correlates with granulocyte differentiation and maturation. However, the physiological role of Ly6G alone remains still unclear. The monoclonal antibody NIMP-R14 has been successfully used to stain polymorphonuclear (PMN) cells and monocytes for fluorescent activated cell sorting and in frozen and paraffin sections. Treatment with antibodies in vivo leads to neutropenia and has inhibitory effect on local immune responses. Furthermore, it has been shown to be useful for depletion of neutrophils in mice. It depletes neutrophils as soon as 6 hours after injection and up to 6 days.
  • Clonality
    Monoclonal
  • Clone number
    NIMP-R14
  • Myeloma
    210RCY3-Ag123
  • Isotype
    IgG2b
  • Light chain type
    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2557 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
ICC Use at an assay dependent concentration.
Functional Studies Use at an assay dependent concentration. Neutrophil depletion. Mice were treated with NIMP-R14 given intraperitoneally at a dose of 1 mg, 6 hours before infection.
IHC-P Use at an assay dependent concentration.

This antibody has been reported to work well without antigen retrieval, or with retrieval in a 125 degree C decloaking chamber for 3 minutes with pH 6.5 buffer. HIER optimization may be required.

Flow Cyt Use at an assay dependent concentration.

5 x 105 cells were incubated with 10 µg/ml antibody.
ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration. Tested with acetone-fixed tissue.

Target

  • Cellular localization
    Cell membrane; Lipid-anchor, GPI-anchor
  • Database links
  • Alternative names
    • Ly-6B antibody
    • Ly6b antibody
    • Lymphocyte antigen 6B antibody
    • Lymphocyte antigen 6B Short name=Ly-6B antibody

Images

  • Frozen sections of mouse spleen. ab2557 was used in a concentration of 5 μg/ml.

  • ab2557 at 1/80 staining mouse blood cells (ip wash cells) by Immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a biotinylated rabbit anti-rat (mouse adsorbed) antibody.

    This image is courtesy of an Abreview submitted on 16 March 2006.

    See Abreview

  • ab2557 staining Neutrophil in Mouse lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formalin and blocked with 2.5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10mM sodium citrate. Samples were incubated with primary antibody (1/200 in 1x PBS/1% BSA/0.3% Triton X-100 buffer) for 1 hour at 25°C. A HRP-conjugated Goat anti-rat Ig polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab2557 staining Ly-6G/-6C in mouse spleen tissue by Immunohistochemistry (Frozen sections).
  • ab2557 staining Neutrophil in mouse tumor tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed. Samples were then blocked, followed by incubation with the primary antibody at a 1/50 dilution for 30 minutes. A biotin-conjugated rabbit anti-rat was used as secondary antibody at a 1/400 dilution.

References

This product has been referenced in:
  • Bodas M  et al. Inhibition of histone-deacetylase activity rescues inflammatory cystic fibrosis lung disease by modulating innate and adaptive immune responses. Respir Res 19:2 (2018). IHC-P ; Mouse . Read more (PubMed: 29301535) »
  • Mahmutovic Persson I  et al. IL-1ß mediates lung neutrophilia and IL-33 expression in a mouse model of viral-induced asthma exacerbation. Respir Res 19:16 (2018). IHC-P ; Mouse . Read more (PubMed: 29361942) »

See all 82 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
None
Permeabilization
No
Specification
brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde
Username

Herr Dr. Markus Kipp

Verified customer

Submitted Sep 07 2018

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 (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH 6.0
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

Submitted May 10 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 18 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM Citrate
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Wesley Konsavage

Verified customer

Submitted Dec 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Victoria Bridgeman

Verified customer

Submitted Jun 23 2016

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)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph6 citrate
Permeabilization
No
Specification
Skin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Victoria Bridgeman

Verified customer

Submitted Jun 23 2016

Application
Flow Cytometry
Fixation
None, cells were used fresh
Permeabilization
No
Sample
Mouse Cell (Cells from bronchoaveolar lavage in balb/c mice)
Specification
Cells from bronchoaveolar lavage in balb/c mice
Gating Strategy
alive (DAPI-),CD45+, C11b+, CD11c- cells
Preparation
Cell harvesting/tissue preparation method: Bronchoaveolar lavage in balb/c mice was performed with PBS-EDTA 3mM
Sample buffer: PBS-EDTA 3mM
Username

Dr. Francois Daubeuf

Verified customer

Submitted Feb 20 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM sodium citrate
Sample
Mouse Tissue sections (lung)
Specification
lung
Permeabilization
No
Fixative
Formalin
Username

Abcam user community

Verified customer

Submitted Jan 02 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Sample
Mouse Tissue sections (brain)
Specification
brain
Permeabilization
Yes - Triton-X
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 05 2013

Thank you for contacting us.

Unfortunately, we cannot test every organ or tissue when we establish a product.However, if the knee joints were decalcified properly and the species is tested, I don't see any problems why these antibodies shoul...

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