Product nameAnti-Basophils antibody [2D7]
DescriptionMouse monoclonal [2D7] to Basophils
Tested applicationsSuitable for: WB, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Human
Lysate of purified Basophils.
- Chronic myeloid leukaemia (basophilia). Negative control: CD203c negative cells (Basophil depleted cell fractions).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: 99% PBS, 0.1% BSA
Concentration information loading...
PurityProtein G purified
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.
|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.|
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.|
RelevanceBasophils 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.
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.
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.
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.
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
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.
This product has been referenced in:
- Hashimoto T et al. Pathophysiological mechanisms of itch in bullous pemphigoid. J Am Acad Dermatol N/A:N/A (2019). Read more (PubMed: 31351883) »
- Yanagawa M et al. Basophils activated via TLR signaling may contribute to pathophysiology of type 1 autoimmune pancreatitis. J Gastroenterol 53:449-460 (2018). IHC-P ; Human . Read more (PubMed: 28921377) »