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      BUV395 Rat Anti-Mouse I-A, I-E
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      This product is the replacement for 743876.
      BUV395 Rat Anti-Mouse I-A, I-E
      Multiparameter flow cytometric analysis of I-A/I-E expression on Mouse splenocytes. C57BL/6 Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD45R/B220 (Cat. No. 553090/ 553089/561878) and PE Rat Anti-CD11b (Cat. No. 553311/557397/561689) antibodies and with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; Left Plot) or BD Horizon™ BUV395 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 569244 Right Plot) at 0.125 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of I-A/I-E (or Ig Isotype control staining) versus CD45R/B220 and CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7AAD-negative) leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BUV395 Rat Anti-Mouse I-A, I-E
      Multiparameter flow cytometric analysis of I-A/I-E expression on Mouse splenocytes. C57BL/6 Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD45R/B220 (Cat. No. 553090/ 553089/561878) and PE Rat Anti-CD11b (Cat. No. 553311/557397/561689) antibodies and with either BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; Left Plot) or BD Horizon™ BUV395 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 569244 Right Plot) at 0.125 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of I-A/I-E (or Ig Isotype control staining) versus CD45R/B220 and CD11b was derived from gated events with the forward and side light-scatter characteristics of viable (7AAD-negative) leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Horizon™
      H-2I; I-Ad, -Ed; I-Eb and I-Ek MHC class II alloantigens; Ia Ag; MHC II
      Mouse (QC Testing)
      Rat DA, also known as DA/HA IgG2a, κ
      BALB/c mouse epidermal Langerhans cells
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      14961, 14969
      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 and unconjugated antibody and free dye were removed.
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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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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      569244 Rev. 1
      Antibody Details
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      2G9

      The 2G9 monoclonal antibody reacts with the I-Ad and I-Ed MHC class II alloantigens.  It also reacts with transfectants expressing I-Eb and I-Ek and with cells from mice of the H-2p and H-2q haplotypes.  This antibody is non-reactive with cells from SJL (H-2s) and NOD (H-2g7) mice.  Flow cytometric analysis indicates that the 2G9 and M5/114.15.2 monoclonal antibodies have comparable reactivity on cells from mice with I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek alloantigens.

      569244 Rev. 1
      Format Details
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      BUV395
      The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV395
      Ultraviolet 355 nm
      348 nm
      395 nm
      569244 Rev.1
      Citations & References
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      View product citations for antibody "569244" on CiteAb

      Development References (3)

      1. Becker D, Mohamadzadeh M, Reske K, Knop J. Increased level of intracellular MHC class II molecules in murine Langerhans cells following in vivo and in vitro administration of contact allergens. J Invest Dermatol. 1992; 99(5):545-549. (Immunogen: Flow cytometry, Fluorescence activated cell sorting, Immunofluorescence, Immunoprecipitation). View Reference
      2. Farquhar CA, Paterson AM, Cobbold SP, et al. Tolerogenicity is not an absolute property of a dendritic cell. Eur J Immunol. 2010; 40(6):1728-1737. (Clone-specific: Flow cytometry). View Reference
      3. Wculek SK, Amores-Iniesta J, Conde-Garrosa R, Khouili SC, Melero I, Sancho D. Effective cancer immunotherapy by natural mouse conventional type-1 dendritic cells bearing dead tumor antigen.. J Immunother Cancer. 2019; 7(1):100. (Clone-specific). View Reference
      569244 Rev. 1

       

      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.