The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50. Fresh surgical tissues can be fixed in Carnoy's fluid for 24 h and transferred to absolute ethanol.
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.
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.
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Basophils antibody [2D7] (ab155577)This image is courtesy of an anonymous Abreview
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.
Flow Cytometry - Anti-Basophils antibody [2D7] (ab155577)This image is courtesy of an anonymous 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.
Western blot - Anti-Basophils antibody [2D7] (ab155577)This image is courtesy of an anonymous 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
Immunohistochemistry (Frozen sections) - Anti-Basophils antibody [2D7] (ab155577)This image is courtesy of an anonymous 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.
Yanagawa M et al. Basophils activated via TLR signaling may contribute to pathophysiology of type 1 autoimmune pancreatitis. J Gastroenterol53:449-460 (2018).
Read more (PubMed: 28921377) »
Kim DK et al. MBP-positive and CD11c-positive cells are associated with different phenotypes of Korean patients with non-asthmatic chronic rhinosinusitis. PLoS One9:e111352 (2014).
Read more (PubMed: 25361058) »