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BV421 Mouse Anti-Human IgE
BV421 Mouse Anti-Human IgE

Two-color flow cytometric analysis of IgE expression on human peripheral blood leucocytes. Whole blood cells were stained with BD Horizon™ PE-CF594 Mouse Anti-Human CD193 antibody (Cat. No. 562571) and either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human IgE antibody (Cat. No. 566324/566325; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot showing the correlated expression of CD193 versus IgE (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Two-color flow cytometric analysis of IgE expression on human peripheral blood leucocytes. Whole blood cells were stained with BD Horizon™ PE-CF594 Mouse Anti-Human CD193 antibody (Cat. No. 562571) and either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; Left Plot) or BD Horizon BV421 Mouse Anti-Human IgE antibody (Cat. No. 566324/566325; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot showing the correlated expression of CD193 versus IgE (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
Immunoglobulin E; IGHE; IGHε; Ig epsilon chain constant region
Human (QC Testing)
Mouse IgG2a, κ
Flow cytometry (Routinely Tested)
5 µl
AB_2744487
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566325 Rev. 1
Antibody Details
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G7-26

The G7-26 monoclonal antibody specifically recognizes the constant region of human Immunoglobulin E (IgE). It does not crossreact with other immunoglobulin heavy chain isotypes. IgE exists in a transmembrane form that is expressed by B lymphocytes and serves as an antigen receptor. Soluble IgE is produced and secreted by activated B cells and plasma cells. IgE may bind through its constant region to cell surface receptors, such as the high-affinity (FcεRI) or low-affinity (FcεRII/CD23) receptors for IgE. FcεRI is expressed on mast cells, basophils, and at lower levels, on dendritic cells and monocytes. FcεRII/CD23 is expressed by B cells and by some other cell types including T cells, monocytes, eosinophils, neutrophils, follicular dendritic cells, and Langerhans cells. Crosslinking of Fc receptor-bound IgE antibodies by multivalent antigens or allergens can induce phagocytosis or the cellular release of inflammatory mediators. Although IgE can provide immune protection against pathogenic parasites, it may also play a central role in a variety of allergic disorders.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

566325 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
566325 Rev.1
Citations & References
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Development References (5)

  1. Böhm I, Speck U, Schild HH. Pilot study on basophil activation induced by contrast medium.. Fundam Clin Pharmacol. 2011; 25(2):267-76. (Clone-specific: Flow cytometry). View Reference
  2. Cheng YX, Foster B, Holland SM, et al. CD2 identifies a monocyte subpopulation with immunoglobulin E-dependent, high-level expression of Fc epsilon RI.. Clin Exp Allergy. 2006; 36(11):1436-45. (Clone-specific). View Reference
  3. Foster B, Metcalfe DD, Prussin C. Human dendritic cell 1 and dendritic cell 2 subsets express FcepsilonRI: correlation with serum IgE and allergic asthma.. J Allergy Clin Immunol. 2003; 112(6):1132-8. (Biology). View Reference
  4. Horst A, Hunzelmann N, Arce S, et al. Detection and characterization of plasma cells in peripheral blood: correlation of IgE+ plasma cell frequency with IgE serum titre.. Clin Exp Immunol. 2002; 130(3):370-8. (Clone-specific: Flow cytometry). View Reference
  5. Kubota T, Mukai K, Minegishi Y, Karasuyama H. Different stabilities of the structurally related receptors for IgE and IgG on the cell surface are determined by length of the stalk region in their alpha-chains.. J Immunol. 2006; 176(11):7008-14. (Clone-specific: Flow cytometry). View Reference
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566325 Rev. 1

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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.