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BUV615 Mouse Anti-Human IgD
BUV615 Mouse Anti-Human IgD

Two-color flow cytometric analysis of IgD expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with PE Mouse Anti-Human CD19 antibody (Cat. No. 555413/561741) and either BD Horizon™ BUV615 Mouse IgG2a, κ Isotype Control (Cat. No. 613009; Left Plot) or BD Horizon BUV615 Mouse Anti-Human IgD antibody (Cat. No. 613008; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of IgD (or Ig isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Two-color flow cytometric analysis of IgD expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were cultured in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with PE Mouse Anti-Human CD19 antibody (Cat. No. 555413/561741) and either BD Horizon™ BUV615 Mouse IgG2a, κ Isotype Control (Cat. No. 613009; Left Plot) or BD Horizon BUV615 Mouse Anti-Human IgD antibody (Cat. No. 613008; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of IgD (or Ig isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Product Details
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BD Horizon™
IGHD; Ig delta chain C region; Immunoglobulin heavy constant delta
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human IgD
Flow cytometry (Routinely Tested)
5 µl
3495
AB_2870276
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

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) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
613008 Rev. 1
Antibody Details
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IA6-2

The IA6-2 monoclonal antibody specifically binds to the heavy chain of human Immunoglobulin D (IgD). IgD is a member of the immunoglobulin superfamily that exists in type 1-membrane (mIgD) and soluble glycoprotein forms. mIgD is expressed on mature naïve B cells (along with membrane IgM) and serves as a B-cell receptor for antigen (BCR).  In response to antigen binding, the mIgD BCR, in association with other signaling molecules including CD79a and CD79b, can transduce activating or tolerizing signals intracellularly into B lymphocytes.

The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP.  Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).

613008 Rev. 1
Format Details
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BUV615
The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV615
Ultraviolet 355 nm
350 nm
615 nm
613008 Rev.1
Citations & References
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Development References (6)

  1. Kruetzmann S, Rosado MM, Weber H, et al. Human immunoglobulin M memory B cells controlling Streptococcus pneumoniae infections are generated in the spleen. J Exp Med. 2003; 197(7):939-945. (Clone-specific: Flow cytometry). View Reference
  2. Kuritani T, Cooper MD. Human B cell differentiation. III. Enhancing effect of monoclonal anti-immunoglobulin D antibody on pokeweed mitogen-induced plasma cell differentiation.. J Immunol. 1982; 129(6):2490-5. (Clone-specific: Functional assay). View Reference
  3. Odendahl M, Jacobi A, Hansen A, et al. Disturbed peripheral B lymphocyte homeostasis in systemic lupus erythematosus. J Immunol. 2000; 165(10):5970-5979. (Clone-specific: Flow cytometry). View Reference
  4. Preud'homme JL, Petit I, Barra A, Morel F, Lecron JC, Lelievre E. Structural and functional properties of membrane and secreted IgD. Mol Immunol. 2000; 37(15):871-887. (Biology). View Reference
  5. Wei C, Anolik J, Cappione A, et al. A new population of cells lacking expression of CD27 represents a notable component of the B cell memory compartment in systemic lupus erythematosus.. J Immunol. 2007; 178(10):6624-33. (Clone-specific: Flow cytometry). View Reference
  6. White MB, Shen AL, Word CJ, Tucker PW, Blattner FR. Human immunoglobulin D: genomic sequence of the delta heavy chain. Science. 1985; 228(4700):733-737. (Biology). View Reference
View All (6) View Less
613008 Rev. 1

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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.