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V500 Mouse Anti-Human IgD
V500 Mouse Anti-Human IgD

Multicolor flow cytometric analysis of IgD expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were incubated in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697) and with either BD Horizon™ V500 Mouse IgG2a, κ Isotype Control (Cat. No. 561221; Left Panel) or BD Horizon™ V500 Mouse anti-Human IgD antibody (Cat. No. 561490; Right Panel). The two-color flow cytometric dot plots showing the correlated expression of IgD (or Ig isotype control staining) versus CD19 were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Multicolor flow cytometric analysis of IgD expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were incubated in complete tissue culture medium overnight in order to minimize subsequent nonspecific immunofluorescent staining. The cells were harvested and stained with Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697) and with either BD Horizon™ V500 Mouse IgG2a, κ Isotype Control (Cat. No. 561221; Left Panel) or BD Horizon™ V500 Mouse anti-Human IgD antibody (Cat. No. 561490; Right Panel). The two-color flow cytometric dot plots showing the correlated expression of IgD (or Ig isotype control staining) versus CD19 were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

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_10679356
Aqueous buffered solution containing protein stabilizer, glycerol and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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. BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561490 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 is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is

available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.

When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls.  Due to spectral differences between cells and beads  in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents.   Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.   Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.

561490 Rev. 1
Format Details
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V500
BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V500
Violet 405 nm
415 nm
499 nm
561490 Rev.1
Citations & References
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Development References (3)

  1. 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
  2. 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
  3. 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
561490 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.