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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
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
Companion Products
The 9B9 monoclonal antibody specifically binds to CD368, which is also known as CLEC6 (C-type lectin-like receptor 6), CLEC4D, CLECSF8, and MCL (macrophage C-type lectin). CD368 is a 30 kDa type II transmembrane glycoprotein that belongs to the C-type Lectin Receptor family of molecules. It is primarily expressed on peripheral blood neutrophils and monocytes and weakly on several subsets of blood dendritic cells. Expression of this receptor is lost upon in vitro differentiation of monocytes into dendritic cells or macrophages. CD368 reportedly serves as an endocytic receptor that may function in antigen clearance and presentation to T lymphocytes. CD368 may also play roles in cellular adhesion, signaling and glycoprotein turnover. CD368 expression can be upregulated by cells in response to cytokines including IFN-γ, TNF, IL-6, and IL-10.
Development References (5)
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Arce I, Martínez-Muñoz L, Roda-Navarro P, Fernández-Ruiz E. The human C-type lectin CLECSF8 is a novel monocyte/macrophage endocytic receptor.. Eur J Immunol. 2004; 34(1):210-20. (Biology). View Reference
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Autenrieth SE, Grimm S, Rittig SM, Grünebach F, Gouttefangeas C, Bühring HJ. Profiling of primary peripheral blood- and monocyte-derived dendritic cells using monoclonal antibodies from the HLDA10 Workshop in Wollongong, Australia.. Clin Transl Immunology. 2015; 4(11):e50. (Clone-specific: Flow cytometry). View Reference
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Graham LM, Gupta V, Schafer G, et al. The C-type lectin receptor CLECSF8 (CLEC4D) is expressed by myeloid cells and triggers cellular activation through Syk kinase.. J Biol Chem. 2012; 287(31):25964-74. (Biology). View Reference
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Gurka S, Dirks S, Photiadis J, Kroczek RA. Expression analysis of surface molecules on human thymic dendritic cells with the 10th HLDA Workshop antibody panel.. Clin Transl Immunology. 2015; 4(10):e47. (Clone-specific). View Reference
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Ohradanova-Repic A, Machacek C, Fischer MB, Stockinger H. Differentiation of human monocytes and derived subsets of macrophages and dendritic cells by the HLDA10 monoclonal antibody panel.. Clin Transl Immunology. 2016; 5(1):e55. (Clone-specific: Flow cytometry). View Reference
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.