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BV421 Goat Anti-Rabbit IgG

BV421 Goat Anti-Rabbit IgG

Clone Polyclonal

(RUO)
BV421 Goat Anti-Rabbit IgG
Immunofluorescent Staining using BD Horizon™ BV421 Goat Anti-Rabbit Ig     Left Panel - Immunofluorescence staining of apoptotic cells. HeLa cells were treated with Camptothecin (20 µM, 6 hr) to induce apoptosis. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized (5 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050), washed with 1× PBS, and blocked (30 min) with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1× PBS. The cells were stained (1 hr) with Purified Mouse Anti-Cytochrome c (Cat. No. 556432) and Purified Rabbit Anti-Active Caspase-3 (Cat. No. 559565) antibodies at 2.5 µg/mL diluted in blocking buffer. After washing, the cells were stained (45 min) with the second step reagents, BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014; pseudo-colored green) and BD Horizon™ BV480 Goat Anti-Mouse Ig (Cat. No. 564877, pseudo-colored red) antibodies at 2.5 µg/mL diluted in blocking buffer. DRAQ5 (Cat. No. 564902/564903; pseudo-colored blue) was used as a nuclear counterstain. Images were acquired with a standard epifluorescence microscope with a 20× objective.     Right Panel - Multiparameter flow cytometric analysis of CD88 expression on human peripheral blood leucocytes. Whole blood was either not stained with a primary antibody as a control (Top Plot) or primarily stained with Purified Rabbit Anti-Human CD88 monoclonal antibody (Cat. No. 559159; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The cells were washed and secondarily stained with BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014). Two-parameter flow cytometric contour plots showing the correlated expression of CD88 (or Unstained control) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Immunofluorescent Staining using BD Horizon™ BV421 Goat Anti-Rabbit Ig     Left Panel - Immunofluorescence staining of apoptotic cells. HeLa cells were treated with Camptothecin (20 µM, 6 hr) to induce apoptosis. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized (5 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050), washed with 1× PBS, and blocked (30 min) with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1× PBS. The cells were stained (1 hr) with Purified Mouse Anti-Cytochrome c (Cat. No. 556432) and Purified Rabbit Anti-Active Caspase-3 (Cat. No. 559565) antibodies at 2.5 µg/mL diluted in blocking buffer. After washing, the cells were stained (45 min) with the second step reagents, BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014; pseudo-colored green) and BD Horizon™ BV480 Goat Anti-Mouse Ig (Cat. No. 564877, pseudo-colored red) antibodies at 2.5 µg/mL diluted in blocking buffer. DRAQ5 (Cat. No. 564902/564903; pseudo-colored blue) was used as a nuclear counterstain. Images were acquired with a standard epifluorescence microscope with a 20× objective.     Right Panel - Multiparameter flow cytometric analysis of CD88 expression on human peripheral blood leucocytes. Whole blood was either not stained with a primary antibody as a control (Top Plot) or primarily stained with Purified Rabbit Anti-Human CD88 monoclonal antibody (Cat. No. 559159; Bottom Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The cells were washed and secondarily stained with BD Horizon™ BV421 Goat Anti-Rabbit Ig (Cat. No. 565014). Two-parameter flow cytometric contour plots showing the correlated expression of CD88 (or Unstained control) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
Rabbit (QC Testing)
Goat Ig
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
AB_2716308
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.

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.
565014 Rev. 1
Antibody Details
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Poly1272

BD Horizon™ BV421 Goat Anti-Rabbit IgG is intended to be a second-step reagent for immunofluorescent staining of cells pre-stained with Rabbit IgG primary antibodies. It reacts with rabbit IgG and the light chains of other rabbit immunoglobulins.  BV421 Anti-Rabbit IgG has minimal crossreactivity with human, mouse, or rat serum proteins. As a second step staining reagent, it is reactive with rabbit polyclonal and monoclonal antibodies.

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.

565014 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.
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BV421
Violet 405 nm
407 nm
423 nm
565014 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.