Overview

  • Product name
    Anti-Basophils antibody [2D7]
  • Description
    Mouse monoclonal [2D7] to Basophils
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Lysate of purified Basophils.

  • Positive control
    • Chronic myeloid leukaemia (basophilia). Negative control: CD203c negative cells (Basophil depleted cell fractions).

Properties

Applications

Our Abpromise guarantee covers the use of ab155577 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
WB 1/50.
IHC-P 1/50. Fresh surgical tissues can be fixed in Carnoy's fluid for 24 h and transferred to absolute ethanol.
IHC-Fr 1/50. Fix in either 10% NBF or Carnoy's fluid for 15 minutes at RT.
Flow Cyt 1/50.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/50. Cytospin preparations should be fixed in methanol for 15 min, incubated in methanol containing 0.6% H2O2, for 30 min to block endogenous peroxidase.

Target

  • Relevance
    Basophils are multifunctional haematopoietic cells that secrete a wide variety of proinflammatory agents upon activation such as histamine and cytokines. Moreover, basophils can induce IgE production by B cells. Under normal conditions basophils are primarily produced in the bone marrow, whereas mature basophils reside in the circulation and can enter tissues at sites of inflammation. Both basophils and mast cells have been implicated in the pathogenesis of allergic inflammation due to the abundant expression of high affinity receptors for IgE. The 2D7 antibody can be used as a sensitive and precise marker for human basophils and does not react with lymphocytes, monocytes, eosinophils, neutrophils or mast cells in immunohistochemistry. Moreover, the antibody is suitable for western blot analysis of the 2D7 antigen.

Images

  • ab155577 staining Basophils in human bone marrow tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 4% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer pH 6.0. Samples were incubated with primary antibody (1/50 in blocking buffer) for 16 hours at 22°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (10µg/ml) was used as the secondary antibody.

    See Abreview

  • ab155577 staining Human basophil granulocytes by Flow Cytometry. The sample was incubated with the primary antibody (1/50 in PBS with human IgG (Fc-block)) for 45 minutes at 4°C. A phycoerythrin-conjugated goat anti-mouse IgG polyclonal (1/100) was used as the secondary antibody. Gating Strategy: Basophils gated with FceR1a-antibody. 1 - isotype control, 2 - CD63, 3 - 2D7.

    See Abreview

  • All lanes : Anti-Basophils antibody [2D7] (ab155577) at 1 µg/ml

    Lanes 1-2 : Human basophils purified by magnetic beads
    Lane 3 : Positive control (chronic myeloid leukemia (basophilia))
    Lanes 4-5 : PBMCs
    Lanes 6-7 : Human basophils purified by countercurrent centrifugal elutriation

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated horse anti-mouse IgG polyclonal at 1/2000 dilution

    Performed under reducing conditions.

    Observed band size: 75 kDa (why is the actual band size different from the predicted?)


    Exposure time: 5 minutes

    See Abreview

  • ab155577 staining Basophils in human nasal tissue of allergic rhinitis patient by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 10% NBF and blocked with 4% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/50 in blocking buffer) for 4 hours at 4°C. A Texas Red®-conjugated goat anti-mouse IgG polyclonal (1/100) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Kim DK  et al. MBP-positive and CD11c-positive cells are associated with different phenotypes of Korean patients with non-asthmatic chronic rhinosinusitis. PLoS One 9:e111352 (2014). IHC ; Human . Read more (PubMed: 25361058) »
  • Agis H  et al. Enumeration and immunohistochemical characterisation of bone marrow basophils in myeloproliferative disorders using the basophil specific monoclonal antibody 2D7. J Clin Pathol 59:396-402 (2006). Read more (PubMed: 16461568) »

See all 4 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Nasal mucosa)
Specification
Nasal mucosa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Mr. Duc Dung Le

Verified customer

Submitted Jun 09 2016

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (5-16% gel)
Sample
Human Cell lysate - whole cell (human basophils (see below - additional data))
Specification
human basophils (see below - additional data)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 13 2014

Application
Flow Cytometry
Fixation
no fixation
Permeabilization
No
Sample
Human Cell (basophil granulocytes)
Specification
basophil granulocytes
Gating Strategy
basophils gated with FceR1a-antiboy
Preparation
Cell harvesting/tissue preparation method: peripheral blood cells, separated by Ficoll density separation, followed by magnetic isolation for basophils
Sample buffer: PBS with human IgG (Fc-block)
Username

Abcam user community

Verified customer

Submitted Oct 10 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Sample
Human Tissue sections (nasal tissue of allergic rhinitis patients after n)
Specification
nasal tissue of allergic rhinitis patients after n
Permeabilization
No
Fixative
10% NBF
Username

Abcam user community

Verified customer

Submitted Oct 09 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
Sample
Human Tissue sections (bone marrow)
Specification
bone marrow
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 09 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Sample
Human Cell (PBMC with a high proportion of basophils)
Specification
PBMC with a high proportion of basophils
Permeabilization
Yes - citrate buffer, pH 6.0
Fixative
Methanol
Username

Abcam user community

Verified customer

Submitted Oct 09 2014

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

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