Preparation And Storage
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® CompBeads 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- 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.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The EBVCS-5 antibody specifically binds to human CD23, the low affinity receptor for human IgE (Fc epsilon RII or FcεRII), which is also known as C-type lectin domain family 4 member J (CLEC4J). CD23 is expressed as a ~45 kDa type II membrane glycoprotein that is present at low density on most normal B lymphocytes and at higher levels on activated B lymphocytes, Epstein-Barr virus (EBV)-transformed lymphoblasts, chronic lymphocytic leukemia (CLL) cells of B-lymphocyte origin, and tonsillar B lymphocytes. CD23 expression is induced on B cells by interleukin-4 (IL-4) and is lost after isotype switching to IgA, IgG, or IgE. The CD23 antigen is not present on immature bone marrow B lymphocytes or on T lymphocytes. CD23 may also be differentially expressed on monocytes, macrophages, eosinophils, platelets, dendritic cells, and Langerhans cells. CD23 can mediate IgE-dependent cytotoxicity and phagocytosis by macrophages and eosinophils. Soluble CD23 (sCD23) can be released by CD23-positive cells as a result of proteolytic cleavage of membrane CD23. Larger fragments of sCD23 (eg, 25-37 kDa) retain their IgE-binding capacity whereas smaller fragments (ie, ≤ 12 kDa) do not. Soluble CD23 may have immunoregulatory effects on the growth and differentiation of B cells and other cell types.
Development References (11)
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Bieber T, Rieger A, Neuchrist C, et al. Induction of Fc epsilon R2/CD23 on human epidermal Langerhans cells by human recombinant interleukin 4 and gamma interferon.. J Exp Med. 1989; 170(1):309-14. (Biology). View Reference
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Capron M, Jouault T, Prin L, et al. Functional study of a monoclonal antibody to IgE Fc receptor (Fc epsilon R2) of eosinophils, platelets, and macrophages.. J Exp Med. 1986; 164(1):72-89. (Biology). View Reference
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Choe J, Kim HS, Armitage RJ, Choi YS. The functional role of B cell antigen receptor stimulation and IL-4 in the generation of human memory B cells from germinal center B cells.. J Immunol. 1997; 159(8):3757-66. (Clone-specific: Flow cytometry). View Reference
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Dugas B, Paul-Eugene N, Cairns J, et al. Leukotriene B4 potentiates the expression and release of Fc epsilon RII/CD23, and proliferation and differentiation of human B lymphocytes induced by IL-4.. J Immunol. 1990; 145(10):3406-11. (Clone-specific: ELISA). View Reference
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Kikutani H, Inui S, Sato R, et al. Molecular structure of human lymphocyte receptor for immunoglobulin E.. Cell. 1986; 47(5):657-65. (Biology). View Reference
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Kikutani H, Suemura M, Owaki H, et al. Fc epsilon receptor, a specific differentiation marker transiently expressed on mature B cells before isotype switching.. J Exp Med. 1986; 164(5):1455-69. (Biology). View Reference
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Nadler LM. B Cell/Leukemia Panel Workshop: summary and comments. In: Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, ed. Leukocyte Typing II: Human B Lymphocytes. New York: Springer-Verlag; 1986:3-43.
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Rieber EP, Rank G, Kohler I, Krauss S. Membrane expression of Fc epsilon RII/CD23 and release of soluble CD23 by follicular dendritic cells. Adv Exp Med Biol. 1993; 329:393-398. (Biology). View Reference
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Sarfati M, Ishihara H, Delespesse G. CD23 Workshop Panel report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:530-533.
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Sugden B, Metzenberg S. Characterization of an antigen whose cell surface expression is induced by infection with Epstein-Barr virus.. J Virol. 1983; 46(3):800-7. (Immunogen: Immunoprecipitation, Radioimmunoassay). View Reference
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Yukawa K, Kikutani H, Owaki H, et al. A B cell-specific differentiation antigen, CD23, is a receptor for IgE (Fc epsilon R) on lymphocytes.. J Immunol. 1987; 138(8):2576-80. (Biology). View Reference
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