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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).
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.
- 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.
- 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
The B3506B4 monoclonal antibody specifically recognizes the Fc region of the human Immunoglobulin A (IgA) subclass known as IgA1. IgA1 is comprised of two identical heavy chains encoded by IGHA1, and two light chains, either Igκ or Igλ, that are linked together by disulfide bonds. The B3506B4 antibody does not crossreact with other human immunoglobulin heavy chain (IgH) subclasses, including the IgA2 subclass. IgA1 is expressed by plasmablasts, plasma cells, and memory B cells as well as by some myeloma or plasmacytoma cells. IgA1 is expressed in either a transmembrane form that serves as an antigen-specific B cell surface receptor or in a soluble secreted form. Monomeric and dimeric forms of IgA1 are found in the serum although the monomeric form predominates. Multimeric forms of IgA1 connected by a J-chain and secretory component are also found in bodily secretions. Secretory IgA is transported across epithelial cells and secreted into the lumen of mucosal tissues including the gastrointestinal and respiratory tracts and is found in human breast milk. IgA1 functions in the agglutination of microbes and can block their adherence to or infection of human cells. It can also bind to CD89 (FcαRI) expressed on neutrophils, monocytes, macrophages, and eosinophils to trigger a variety of immune responses including phagocytosis, antibody-dependent cell-mediated cytotoxicity, and the release of inflammatory cytokines and mediators such as in the response to microbes.
Development References (9)
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Blanco E, Perez-Andres M, Sanoja-Flores L, et al. Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells.. J Immunol Methods. 2019; 475:112372. (Clone-specific: Flow cytometry). View Reference
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Blanco E, Pérez-Andrés M, Arriba-Méndez S, et al. Age-associated distribution of normal B-cell and plasma cell subsets in peripheral blood.. J Allergy Clin Immunol. 2018; 141(6):2208-2219.e16. (Clone-specific: Flow cytometry). View Reference
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Delacroix DL, Van Snick J, Vaerman JP, Conley ME, Mascart-Lemone F, Bernier GM. Monoclonal antibodies against isotypic and isoallotypic determinants of human IgA1 and IgA2: fine specificities and binding properties.. Mol Immunol. 1986; 23(4):367-75. (Immunogen: Radioimmunoassay). View Reference
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Fayette J, Dubois B, Vandenabeele S, et al. Human dendritic cells skew isotype switching of CD40-activated naive B cells towards IgA1 and IgA2.. J Exp Med. 1997; 185(11):1909-18. (Biology). View Reference
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He B, Xu W, Santini PA, et al. Intestinal bacteria trigger T cell-independent immunoglobulin A(2) class switching by inducing epithelial-cell secretion of the cytokine APRIL.. Immunity. 2007; 26(6):812-26. (Clone-specific: Flow cytometry). View Reference
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Sibille Y, Chatelain B, Staquet P, Merrill WW, Delacroix DL, Vaerman JP. Surface IgA and Fc-alpha receptors on human alveolar macrophages from normal subjects and from patients with sarcoidosis.. Am Rev Respir Dis. 1989; 139(3):740-7. (Clone-specific: Blocking). View Reference
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Tangye SG, Ferguson A, Avery DT, Ma CS, Hodgkin PD. Isotype switching by human B cells is division-associated and regulated by cytokines.. J Immunol. 2002; 169(8):4298-306. (Biology). View Reference
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Zan H, Cerutti A, Dramitinos P, Schaffer A, Casali P. CD40 engagement triggers switching to IgA1 and IgA2 in human B cells through induction of endogenous TGF-beta: evidence for TGF-beta but not IL-10-dependent direct S mu-->S alpha and sequential S mu-->S gamma, S gamma-->S alpha DNA recombination.. J Immunol. 1998; 161(10):5217-25. (Biology). View Reference
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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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.