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      V500 Rat Anti-Mouse I-A/I-E
      V500 Rat Anti-Mouse I-A/I-E
      Multicolor flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on splenocytes from positive and negative mouse strains. Mouse spleen cells from either M5/114-negative SJL (Left Panel) or M5/114-positive BALB/c (Right Panel) mice were stained with BD Horizon™ V500 Rat Anti-Mouse  I-A/I-E (Cat. No. 562366), APC Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and APC Anti-Mouse CD11b (Cat. No. 553312/561690) antibodies. Two-color flow cytometric dot plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. The M5/114 monoclonal antibody detects I-Ab,d,q, and I-Ed,k MHC class II alloantigens that are expressed on both B cells and macrophages. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      V500 Rat Anti-Mouse I-A/I-E
      Multicolor flow cytometric analysis of I-A/I-E MHC class II alloantigen expression on splenocytes from positive and negative mouse strains. Mouse spleen cells from either M5/114-negative SJL (Left Panel) or M5/114-positive BALB/c (Right Panel) mice were stained with BD Horizon™ V500 Rat Anti-Mouse  I-A/I-E (Cat. No. 562366), APC Anti-Mouse CD45R/B220 (Cat. No. 553092/561880) and APC Anti-Mouse CD11b (Cat. No. 553312/561690) antibodies. Two-color flow cytometric dot plots showing the expression of I-A/I-E MHC class II alloantigens versus CD45R/B220 and CD11b were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. The M5/114 monoclonal antibody detects I-Ab,d,q, and I-Ed,k MHC class II alloantigens that are expressed on both B cells and macrophages. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      H-2I; I-Ab, I-Ad, I-Aq, I-Ed, and I-Ek MHC class II alloAgs; Ia Ag; M5/114; MHC II
      Mouse (QC Testing)
      Rat BN x LEW IgG2b, κ
      Activated C57BL/6 Mouse Spleen Cells
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      111861
      AB_11153488
      Aqueous buffered solution containing protein stabilizer, glycerol 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™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      6. An isotype control should be used at the same concentration as the antibody of interest.
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      562366 Rev. 2
      Antibody Details
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      M5/114.15.2

      The M5/114.15.2 monoclonal antibody recognizes a polymorphic determinant shared by the I-A[b], I-A[d], I-A[q], I-E[d], and I-E[k] (but not I-A[f], I-A[k], or I-A[s]) MHC class II alloantigens that can be expressed by B cells, dendritic cells, monocytes, macrophages and activated T cells. It also reacts with cells from mice of the H-2[p] and H-2[r] haplotypes, and it is non-reactive with cells from NOD (H-2[g7]) mice. Flow cytometric analysis indicates that the M5/114.15.2 and 2G9 monoclonal antibodies have comparable reactivity on cells from mice with I-A[b], I-A[d], I-A[g7], I-A[q], I-E[d], and I-E[k] alloantigens.

      The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is

      available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.

      When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls.  Due to spectral differences between cells and beads  in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents.   Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.   Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.

      562366 Rev. 2
      Format Details
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      V500
      BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      V500
      Violet 405 nm
      415 nm
      499 nm
      562366 Rev.2
      Citations & References
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      View product citations for antibody "562366" on CiteAb

      Development References (7)

      1. Bhattacharya A, Dorf ME, Springer TA. A shared alloantigenic determinant on Ia antigens encoded by the I-A and I-E subregions: evidence for I region gene duplication. J Immunol. 1981; 127(6):2488-2495. (Immunogen: Immunoprecipitation). View Reference
      2. Ernst DN, McQuitty DN, Weigle WO, Hobbs MV. Expression of membrane activation antigens on murine B lymphocytes stimulated with lipopolysaccharide. Cell Immunol. 1988; 114(1):161-173. (Clone-specific: Flow cytometry). View Reference
      3. Guo MW, Watanabe T, Mori E, Mori T. Molecular structure and function of CD4 on murine egg plasma membrane. Zygote. 1995; 3(1):65-73. (Clone-specific: Blocking). View Reference
      4. Hattori M, Buse JB, Jackson RA, et al. The NOD mouse: recessive diabetogenic gene in the major histocompatibility complex. Science. 1986; 231(4739):733-735. (Clone-specific). View Reference
      5. Nelson AJ, Hosier S, Brady W, Linsley PS, Farr AG. Medullary thymic epithelium expresses a ligand for CTLA4 in situ and in vitro. J Immunol. 1998; 151(5):2453-2461. (Clone-specific: Blocking, Immunofluorescence, Immunohistochemistry). View Reference
      6. Viville S, Neefjes J, Lotteau V, et al. Mice lacking the MHC class II-associated invariant chain. Cell. 1993; 72(4):635-648. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
      7. Yamashita I, Nagata T, Tada T, Nakayama T. CD69 cell surface expression identifies developing thymocytes which audition for T cell antigen receptor-mediated positive selection. Int Immunol. 1993; 5(9):1139-1150. (Clone-specific: Blocking). View Reference
      View All (7) View Less
      562366 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.