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PE Mouse Anti-Human Basophils (2D7)
PE Mouse Anti-Human Basophils (2D7)
Multiparameter flow cytometric analysis of Basophils (2D7) expression on human peripheral blood leucocyte populations. Human whole blood was fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and permeabilized with BD Phosflow™  Perm/Wash Buffer I (Cat. No. 557885), and the cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Basophils (2D7)antibody (Cat. No. 568076/568077; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of Basophils (2D7) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
PE Mouse Anti-Human Basophils (2D7)

Two-color flow cytometric analysis of Basophils (2D7) expression on human peripheral blood leucocytes. Human whole blood was stained with BV421 Mouse Anti-Human CD193 antibody (Cat. No. 562570) and then fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and permeabilized with BD Phosflow™  Perm/Wash Buffer I (Cat. No. 557885). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Basophils (2D7)  antibody (Cat. No. 568076/568077; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of Basophils (2D7) [or Ig Isotype control staining] versus CD193 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Multiparameter flow cytometric analysis of Basophils (2D7) expression on human peripheral blood leucocyte populations. Human whole blood was fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and permeabilized with BD Phosflow™  Perm/Wash Buffer I (Cat. No. 557885), and the cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Basophils (2D7)antibody (Cat. No. 568076/568077; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of Basophils (2D7) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Two-color flow cytometric analysis of Basophils (2D7) expression on human peripheral blood leucocytes. Human whole blood was stained with BV421 Mouse Anti-Human CD193 antibody (Cat. No. 562570) and then fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) and permeabilized with BD Phosflow™  Perm/Wash Buffer I (Cat. No. 557885). The cells were then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Human Basophils (2D7)  antibody (Cat. No. 568076/568077; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of Basophils (2D7) [or Ig Isotype control staining] versus CD193 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Pharmingen™
Basophils; Basophils (2D7); 2D7 Antigen; 2D7 Ligand
Human (QC Testing)
Mouse BALB/c IgG1, κ
Purified Human Basophil Lysate
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Antibody Details
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2D7/BASO

The 2D7 monoclonal antibody selectively recognizes human basophils, Basophils (2D7). The antibody binds to a ~7.2 to 7.5 kDa basophil-specific protein, also known as the 2D7 antigen or ligand, present in secretory granules which has not been conclusively identified. Basophils normally comprise a relatively small percentage (<1%) of total peripheral blood leucocytes. Basophils express high levels of FceRI, a high affinity receptor for IgE. Upon activation through crosslinking of FceRI, basophils degranulate and may thus show reduced staining with the 2D7 antibody due to the release or destruction of the 2D7 antigen.  The activated basophils release proinflammatory mediators that can either counter parasitic infections or lead to acute or chronic allergic diseases, including anaphylaxis, asthma, and atopic dermatitis. Activated basophils release a variety of mediators including the vasodilator histamine, proteolytic enzymes, and several cytokines which promote inflammation as well as heparin which  prevents blood clotting. The 2D7 monoclonal antibody is useful for identifying human basophils since it does not recognize lymphocytes, monocytes, eosinophils, neutrophils, or mast cells.

        

Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
Citations & References
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Development References (6)

  1. Agis H, Krauth MT, Mosberger I, et al. Enumeration and immunohistochemical characterisation of bone marrow basophils in myeloproliferative disorders using the basophil specific monoclonal antibody 2D7. J Clin Pathol. 2006; 59(4):396-402. (Clone-specific: Immunohistochemistry). View Reference
  2. Foster B, Schwartz LB, Devouassoux G, Metcalfe DD, Prussin C. Characterization of mast-cell tryptase-expressing peripheral blood cells as basophils. J Allergy Clin Immunol. 2002; 109(2):287-293. (Clone-specific: Flow cytometry). View Reference
  3. Horny HP, Sotlar K, Stellmacher F, et al. The tryptase positive compact round cell infiltrate of the bone marrow (TROCI-BM): a novel histopathological finding requiring the application of lineage specific markers. J Clin Pathol. 2006; 59(3):298-302. (Clone-specific: Immunohistochemistry). View Reference
  4. Karasuyama H, Obata K, Wada T, Tsujimura Y, Mukai K. Newly appreciated roles for basophils in allergy and protective immunity.. Allergy. 2011; 66(9):1133-41. (Clone-specific). View Reference
  5. Kepley CL, Craig SS, Schwartz LB. Identification and partial characterization of a unique marker for human basophils.. J Immunol. 1995; 154(12):6548-55. (Immunogen: Electron microscopy, Immunocytochemistry, Western blot). View Reference
  6. Plager DA, Weiss EA, Kephart GM, et al. Identification of basophils by a mAb directed against pro-major basic protein 1. J Allergy Clin Immunol. 2006; 117(3):626-634. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.