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    BV421 Goat Anti-Rat Ig

    BD Horizon™ BV421 Goat Anti-Rat Ig

    Clone Polyclonal

    (RUO)
    BV421 Goat Anti-Rat Ig
    Immunofluorescent Staining using BD Horizon™ BV421 Goat Anti-Rat Ig.      Left Panel: Multicolor image analysis of CD3e+ CD4+ cells within mouse spleen. Mouse spleen cryosections (5 μm) were 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-Mouse CD4 antibody (Cat. No. 553043) at 2.5 μg/mL. After washing, the sections were secondarily stained with BD Horizon BV421 Goat Anti-Rat Ig (Cat. No. 565013; pseudo-colored green; Upper Right Image) at 2.5 μg/mL. Sections were then washed and counterstained with Alexa Fluor® 488 Hamster Anti-Mouse CD3e antibody (Cat. No. 557666; pseudo-colored red; Upper Left) and DRAQ5™; pseudo-colored blue; Lower Left) as a nuclear stain. Slides were mounted with ProLong® Gold and the images were captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software (Lower Right). 20X objective.      Right Panel: Flow cytometric analysis of CD8b.2 or CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were either not stained with a primary antibody (control staining; dashed line histogram) or stained with Purified Rat Anti-Mouse CD8b.2 antibody (Cat. No. 553038; solid line histogram; Upper Plot) or Purified Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553084/557390; solid line histogram; Lower Plot), as indicated. The cells were washed and secondarily stained with BD Horizon BV421 Goat Anti-Rat Ig. Fluorescence histograms showing either CD8b.2 or CD45R/B220 expression (or control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer.
    BV421 Goat Anti-Rat Ig
    Immunofluorescent Staining using BD Horizon™ BV421 Goat Anti-Rat Ig.      Left Panel: Multicolor image analysis of CD3e+ CD4+ cells within mouse spleen. Mouse spleen cryosections (5 μm) were 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-Mouse CD4 antibody (Cat. No. 553043) at 2.5 μg/mL. After washing, the sections were secondarily stained with BD Horizon BV421 Goat Anti-Rat Ig (Cat. No. 565013; pseudo-colored green; Upper Right Image) at 2.5 μg/mL. Sections were then washed and counterstained with Alexa Fluor® 488 Hamster Anti-Mouse CD3e antibody (Cat. No. 557666; pseudo-colored red; Upper Left) and DRAQ5™; pseudo-colored blue; Lower Left) as a nuclear stain. Slides were mounted with ProLong® Gold and the images were captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software (Lower Right). 20X objective.      Right Panel: Flow cytometric analysis of CD8b.2 or CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were either not stained with a primary antibody (control staining; dashed line histogram) or stained with Purified Rat Anti-Mouse CD8b.2 antibody (Cat. No. 553038; solid line histogram; Upper Plot) or Purified Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553084/557390; solid line histogram; Lower Plot), as indicated. The cells were washed and secondarily stained with BD Horizon BV421 Goat Anti-Rat Ig. Fluorescence histograms showing either CD8b.2 or CD45R/B220 expression (or control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer.
    Product Details
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    BD Horizon™
    Rat (QC Testing)
    Goat Ig
    Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
    0.2 mg/ml
    AB_2739042
    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 monoclonal antibody was purified from tissue culture supernatant or ascites by 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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    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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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    565013 Rev. 1
    Antibody Details
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    Poly1271

    BD Horizon™ BV421 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 crossreactivity detectable by flow cytometry with some, but not all, hamster immunoglobulins.

    The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. BD Horizon BV421 has an Ex Max at 407 nm and Em Max at 421 nm. The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV421.

    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, the recommended excitation and emission filters are 392/23 nm and 430/24 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 flow cytometry, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter).

    565013 Rev. 1
    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
    565013 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.

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