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      BV480 Goat Anti-Rat Ig
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      This SKU will be discontinuing Apr 2024. Suggested alternate SKU is [564877] or for additional support, contact your local applications specialist. Contact Us #

      BD Horizon™ BV480 Goat Anti-Rat Ig

      Clone Polyclonal

      (RUO)
      BV480 Goat Anti-Rat Ig
      Immunofluorescence staining of human LS174T cells. Colorectal adenocarcinoma LS174T cells transfected with human LGR5 (cells from Dr. Hans Clevers, Hubrecht Institute) were cultured and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% Goat serum and 1% BSA diluted in 1x PBS, and stained with Purified Rat Anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562732) at 2.5 μg/ml. After washing, the cells were stained with the second step reagent, BD Horizon™ BV480 Goat Anti-Rat Ig (Cat. No. 564878) (pseudo-colored green) at 2.5 μg/mL. Cells were then washed and permeabilized with 0.1% Triton™ diluted in 1x PBS. Cells were then stained with Alexa Fluor® 488 Mouse Anti-Human Ki-67 antibody (Cat No. 558616) (pseudo-colored red) in blocking buffer with 5% Goat serum, 1% BSA, and 0.5% Triton™ diluted in 1x PBS. DAPI (Cat. No. 564907) was used as a nuclear counterstain (pseudo-colored blue). Slides were mounted with ProLong® Gold and the image was captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software. 40X objective.
      BV480 Goat Anti-Rat Ig
      Immunofluorescence staining of human LS174T cells. Colorectal adenocarcinoma LS174T cells transfected with human LGR5 (cells from Dr. Hans Clevers, Hubrecht Institute) were cultured and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% Goat serum and 1% BSA diluted in 1x PBS, and stained with Purified Rat Anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562732) at 2.5 μg/ml. After washing, the cells were stained with the second step reagent, BD Horizon™ BV480 Goat Anti-Rat Ig (Cat. No. 564878) (pseudo-colored green) at 2.5 μg/mL. Cells were then washed and permeabilized with 0.1% Triton™ diluted in 1x PBS. Cells were then stained with Alexa Fluor® 488 Mouse Anti-Human Ki-67 antibody (Cat No. 558616) (pseudo-colored red) in blocking buffer with 5% Goat serum, 1% BSA, and 0.5% Triton™ diluted in 1x PBS. DAPI (Cat. No. 564907) was used as a nuclear counterstain (pseudo-colored blue). Slides were mounted with ProLong® Gold and the image was captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software. 40X objective.
      Product Details
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      BD Horizon™
      Rat (QC Testing)
      Goat Ig
      Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
      0.2 mg/ml
      AB_2738996
      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 BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 were removed.
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      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. 563794/566349).

      For Immunofluorescence Applications:

      The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480.  For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.  

      For epifluorescence microscopes with broad spectrum excitation sources,  the recommended excitation and emission filters are 445/20 nm and 485/20 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.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
      5. Triton is a trademark of the Dow Chemical Company.
      6. ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
      7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. 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.
      10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564878 Rev. 3
      Antibody Details
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      Poly1271

        BD Horizon™ BV480 Goat Anti-Rat Ig is intended to be a second-step reagent for immunofluorescent staining of cells pre-stained with Rat Ig primary antibodies. It also stains rat B cells and has little reactivity with rat non-B cells or mouse splenocytes. As a second step, it is reactive with rat IgG and IgM monoclonal antibodies although a weaker signal may be detected when the primary antibody has a rat IgM or IgG2b isotype due to its adsorption with mouse Ig. It has weak cross-reactivity detectable by flow cytometry with some, but not all, hamster immunoglobulins.

      The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

      564878 Rev. 3
      Format Details
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      BV480
      The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV480
      Violet 405 nm
      440 nm
      479 nm
      564878 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.

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