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BV480 Goat Anti-Mouse Ig

BV480 Goat Anti-Mouse Ig

Clone Polyclonal

(RUO)
BV480 Goat Anti-Mouse Ig

Immunofluorescence staining of human U-2 OS cells. Cultured cells from the human U-2 OS (Osteosarcoma, ATCC HTB-96) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100, blocked with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1x PBS, and stained with Purified Mouse Anti-GM130 antibody (Cat. No. 610822/610823) at 1.25 μg/mL. After washing, the cells were stained with the second step reagent, Goat BD Horizon™ BV480 Anti-Mouse Ig (Cat. No. 564877) (pseudo-colored green) at 2.5 μg/mL. Cells were then stained with Alexa Fluor® 488 Mouse Anti-β-Tubulin (Cat No. 558605) (pseudo-colored red). 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. 20X objective.

Immunofluorescence staining of human U-2 OS cells. Cultured cells from the human U-2 OS (Osteosarcoma, ATCC HTB-96) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100, blocked with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1x PBS, and stained with Purified Mouse Anti-GM130 antibody (Cat. No. 610822/610823) at 1.25 μg/mL. After washing, the cells were stained with the second step reagent, Goat BD Horizon™ BV480 Anti-Mouse Ig (Cat. No. 564877) (pseudo-colored green) at 2.5 μg/mL. Cells were then stained with Alexa Fluor® 488 Mouse Anti-β-Tubulin (Cat No. 558605) (pseudo-colored red). 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. 20X objective.

Product Details
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BD Horizon™
Mouse (QC Testing)
Goat Ig
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
AB_2738995
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.

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.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. 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.
  9. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  10. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564877 Rev. 3
Antibody Details
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Poly1270

BD Horizon™ BV480 Goat Anti-Mouse Ig is intended to be a second-step reagent for immunofluorescent staining of cells pre-stained with Mouse Ig primary 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 cells occurs in the absence of a primary mouse antibody. In addition, the BD Horizon BV480 Goat Anti-Mouse Ig stains mouse B lymphocytes with little non-specific staining of other cells. Therefore, it is useful as a primary immunofluorescent reagent for staining mouse B cells and antibody-producing cells and as a secondary reagent for staining human and rat cells pre-stained with mouse Ig primary antibodies.

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.

564877 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
564877 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.