Two-color 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 BD Horizon™BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440/562441) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plot) or BD Horizon™ R718 Mouse Anti-Human IgD antibody (Cat. No. 567993/567994; Right Plot) at 1 µg/test. 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 cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
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Two-color 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 BD Horizon™BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440/562441) and with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plot) or BD Horizon™ R718 Mouse Anti-Human IgD antibody (Cat. No. 567993/567994; Right Plot) at 1 µg/test. 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 cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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
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 unreacted dye was 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.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Cell Viability Solution RUO
Size
500 Tests
Cat No.
555815
R718 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
566949
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BV421 Mouse Anti-Human CD19 RUO
Size
100 Tests
Cat No.
562440
567994 Rev. 1
Antibody Details
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.
567994 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
567994 Rev.1
Citations & References
View product citations for antibody "567994" on CiteAb
Development References (5)
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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
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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
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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
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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
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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
567994 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.