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      BV421 Mouse Anti-Human IgG3
      BV421 Mouse Anti-Human IgG3
      Flow cytometric analysis of IgG3 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were washed and cultured in complete tissue culture medium overnight to minimize nonspecific immunofluorescent staining. The cells were harvested and stained with APC Mouse anti-Human CD19 antibody (Cat. No. 555415) and with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD OptiBuild BV421™ Mouse anti-Human IgG3 antibody (Cat. No. 753733; Right Plot) at 0.25 µg per test. The bivariate pseudocolor density plot showing the correlated expression of cell surface IgG3 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BV421 Mouse Anti-Human IgG3
      Flow cytometric analysis of IgG3 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were washed and cultured in complete tissue culture medium overnight to minimize nonspecific immunofluorescent staining. The cells were harvested and stained with APC Mouse anti-Human CD19 antibody (Cat. No. 555415) and with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD OptiBuild BV421™ Mouse anti-Human IgG3 antibody (Cat. No. 753733; Right Plot) at 0.25 µg per test. The bivariate pseudocolor density plot showing the correlated expression of cell surface IgG3 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD OptiBuild™
      IGHG3; Ig gamma-3 chain C region; Immunoglobulin heavy constant gamma 3
      Human (Tested in Development)
      Mouse BALB/c IgG1, κ
      Human IgG3 myeloma protein
      Flow cytometry (Qualified)
      0.2 mg/ml
      3502
      Aqueous buffered solution containing ≤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 that minimize unconjugated dye and antibody.
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      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.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
      6. Researchers should determine the optimal concentration of this reagent for their individual applications.
      7. 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.
      8. 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.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Pacific Blue™ is a trademark of Life Technologies Corporation.
      11. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      12. For U.S. patents that may apply, see bd.com/patents.
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      753733 Rev. 2
      Antibody Details
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      HP6050

      The HP6050 monoclonal antibody specifically recognizes the subclass of human Immunoglobulin G (IgG) known as IgG3. Human IgG3 is the third most abundant of the four subclasses of IgG (IgG1, IgG2, IgG3, and IgG4) named in order of their relative serum concentrations. IgG3 is comprised of two identical heavy chains encoded by IGHG3, and two light chains, either Igκ or Igλ, linked by disulfide bonds. The HP6050 antibody binds to the hinge region of the IgG3 heavy chain and does not crossreact with other immunoglobulin heavy chain (IgH) subclasses. IgG3 is normally expressed by plasmablasts, plasma cells, and memory (IgG3 class-switched) B cells as well as by some myeloma or plasmacytoma cells. HP6050 binds to soluble human IgG3, cytophilic IgG3 attached to Fc receptor-positive cells through its Fc region, or human IgG3 antibodies specifically bound to antigens. IgG3 can cross the placenta and diffuse throughout extravascular fluids throughout the body. Human IgG3 serves multiple functions with the transmembrane form serving as an antigen receptor for B lymphocytes and secreted soluble forms participating in various effector functions. The latter include antibody-dependent neutralization of toxins or infection by microbes, opsonization for phagocytosis, efficient complement fixation, and cell-mediated cytotoxicity.

      753733 Rev. 2
      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.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      753733 Rev.2
      Citations & References
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      View product citations for antibody "753733" on CiteAb

      Development References (6)

      1. Avery DT, Bryant VL, Ma CS, de Waal Malefyt R, Tangye SG. IL-21-induced isotype switching to IgG and IgA by human naive B cells is differentially regulated by IL-4.. J Immunol. 2008; 181(3):1767-79. (Biology). View Reference
      2. Blanco E, Perez-Andres M, Sanoja-Flores L, et al. Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells.. J Immunol Methods. 2019; 475:112372. (Clone-specific: Flow cytometry). View Reference
      3. Gao B, Moore C, Porcheray F, et al. Pretransplant IgG reactivity to apoptotic cells correlates with late kidney allograft loss.. Am J Transplant. 2014; 14(7):1581-91. (Clone-specific: Flow cytometry). View Reference
      4. Jefferis R, Reimer CB, Skvaril F, et al. Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: results of an IUIS/WHO collaborative study.. Immunol Lett. 1985; 10(3-4):223-52. (Clone-specific: ELISA, Hemagglutination assay, Immunocytochemistry, Immunofluorescence, Radioimmunoassay). View Reference
      5. Reimer CB, Phillips DJ, Aloisio CH, et al. Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes.. Hybridoma. 1984; 3(3):263-75. (Immunogen: Array, Fluorescence quantitation). View Reference
      6. Tangye SG, Ferguson A, Avery DT, Ma CS, Hodgkin PD. Isotype switching by human B cells is division-associated and regulated by cytokines.. J Immunol. 2002; 169(8):4298-306. (Biology). View Reference
      View All (6) View Less
      753733 Rev. 2

      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.