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      BV421 Goat Anti-Mouse Ig

      BD Horizon™ BV421 Goat Anti-Mouse Ig

      Clone Polyclonal

      (RUO)
      BV421 Goat Anti-Mouse Ig
      Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes and CD4 expression on rat splenic leucocytes.   Far Left: Human whole blood was incubated with either no primary antibody (dashed line histogram) or Purified Mouse Anti-Human CD3 antibody (Cat. No. 555330; solid line histogram). Human erythrocytes were lysed with BD PharmLyse™ (Cat. No. 555899). Middle Left: Rat splenic leucocytes were similarly incubated with either no antibody (dashed line histogram) or Purified Mouse Anti-Rat CD4 antibody (Cat. No. 554835; solid line histogram).  Both human and rat cells were washed and stained with BD Horizon BV421 Goat Anti-Mouse Ig antibody (Cat. No. 563846). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes or rat leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. Immunofluorescent and immunohistochemical analysis of E-cadherin expression on human breast adenocarcinoma and large intestine. Middle Right: MCF-7 (ATCC HTB-22) were fixed with BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), blocked, and stained with Purified Mouse anti-E-cadherin (Cat. No. 610182) at 5 μg/mL. Second step reagent was BD Horizon BV421 Goat Anti-Mouse Ig at 1.25 μg/mL (pseudo-colored green), captured on a Zeiss LSM 710 confocal microscope, merged using Image J Software. Far Right: Frozen sections of intestine were fixed with BD Cytofix™ buffer, permeabilized with 0.2 % Triton™ X-100, blocked, and stained with Purified Mouse anti-E-cadherin at 5 μg/mL. Second step reagent was BD Horizon BV421 Goat Anti-Mouse Ig at 1.25 μg/mL (pseudo-colored green), captured on a BD Pathway™ 435 Cell Analyzer, merged using BD Attovision™ Software. For both samples, nuclei were counter-stained with DRAQ5™ (pseudo-colored red) and slides were mounted with ProLong® Gold.
      BV421 Goat Anti-Mouse Ig
      Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes and CD4 expression on rat splenic leucocytes.   Far Left: Human whole blood was incubated with either no primary antibody (dashed line histogram) or Purified Mouse Anti-Human CD3 antibody (Cat. No. 555330; solid line histogram). Human erythrocytes were lysed with BD PharmLyse™ (Cat. No. 555899). Middle Left: Rat splenic leucocytes were similarly incubated with either no antibody (dashed line histogram) or Purified Mouse Anti-Rat CD4 antibody (Cat. No. 554835; solid line histogram).  Both human and rat cells were washed and stained with BD Horizon BV421 Goat Anti-Mouse Ig antibody (Cat. No. 563846). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes or rat leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. Immunofluorescent and immunohistochemical analysis of E-cadherin expression on human breast adenocarcinoma and large intestine. Middle Right: MCF-7 (ATCC HTB-22) were fixed with BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), blocked, and stained with Purified Mouse anti-E-cadherin (Cat. No. 610182) at 5 μg/mL. Second step reagent was BD Horizon BV421 Goat Anti-Mouse Ig at 1.25 μg/mL (pseudo-colored green), captured on a Zeiss LSM 710 confocal microscope, merged using Image J Software. Far Right: Frozen sections of intestine were fixed with BD Cytofix™ buffer, permeabilized with 0.2 % Triton™ X-100, blocked, and stained with Purified Mouse anti-E-cadherin at 5 μg/mL. Second step reagent was BD Horizon BV421 Goat Anti-Mouse Ig at 1.25 μg/mL (pseudo-colored green), captured on a BD Pathway™ 435 Cell Analyzer, merged using BD Attovision™ Software. For both samples, nuclei were counter-stained with DRAQ5™ (pseudo-colored red) and slides were mounted with ProLong® Gold.
      Product Details
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      BD Horizon™
      Mouse (QC Testing)
      Goat Ig
      Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry (Tested During Development)
      0.2 mg/ml
      AB_2738449
      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 polyclonal antibody was purified from antiserum by negative adsorption and affinity chromatography. The antibody was conjugated with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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      Recommended Assay Procedures

      For confocal microscopy systems, a 405 nm laser is the optimal excitation source with optimal emission collection centered at 421 nm.

      For epifluorescence microscopes with broad spectrum excitation sources, BD Horizon BV421 can be visualized using standard configurations for

      DAPI or Pacific Blue. The recommended optimal excitation and emission filters are 392/23 nm and 430/25 nm bandpass filters, respectively. For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging

      For imaging applications suboptimal staining can sometimes occur on some nuclear targets.

      For immunofluorescence applications, suboptimal staining can sometimes occur with targets that localize to the nucleus or sub-nuclear compartments, such as Ki-67. In multicolor immunofluorescence applications, we recommend the use of BD Horizon™ BV480 for targets localized to the nucleus and the use of BV421 for cell surface and cytoplasmic targets.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      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. DRAQ5™ is a registered trademark of BioStatus Ltd.
      7. Triton is a trademark of the Dow Chemical Company.
      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.
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      563846 Rev. 3
      Antibody Details
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      Poly1270

      BD Horizon™ BV421 Goat Anti-Mouse Ig is intended to be a second-step reagent for staining of cells pre-stained with primary mouse Ig antibodies. It is reactive with the heavy and light chains of mouse primary antibodies having the IgG1, IgG2a, IgG2b, IgG3, IgM, and IgA isotypes. Minimal background staining of human and rat leucocytes occurs in the absence of a primary mouse antibody. In addition, the BD Horizon BV421 Goat Anti-Mouse Ig stains mouse peripheral B lymphocytes with little non-specific staining of other splenic leucocytes. Therefore, it is useful as a primary reagent in immunofluorescent staining of mouse B cells and antibody-producing cells and as a secondary reagent for staining cells pre-stained with mouse Ig primary antibodies.

      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.

      563846 Rev. 3
      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
      563846 Rev.3
      Citations & References
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      View product citations for antibody "563846" on CiteAb
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      563846 Rev. 3

      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.