BV421 Goat Anti-Mouse Ig
Clone Polyclonal (RUO)
- Brand BD Horizon™
- Concentration 0.2 mg/ml
- Isotype Goat Ig
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
Immunofluorescence, Immunohistochemistry, Bioimaging (Tested During Development)
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
BV421 is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.
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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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- DRAQ5™ is a registered trademark of BioStatus Ltd.
- Triton is a trademark of the Dow Chemical Company.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
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.