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.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The TY/11.8 monoclonal antibody specifically recognizes mouse CD73 which is also known as Ecto-5'-nucleotidase (5'-NT). CD73 is a ~69 kDa glycosylphosphatidylinositol (GPI)-anchored, cell-surface glycoprotein that is encoded by Nt5e which belongs to the 5'-nucleosidase family. CD73 expression appears to be developmentally regulated on leucocytes. In the bone marrow, it is found on most CD11b+ myeloid cells and very few CD19+ cells of the B-lymphocyte lineage. It is neither found expressed on CD11b+ cells in the periphery nor on bone marrow-derived GM-CSF-stimulated dendritic cells. Some peripheral B lymphocytes express CD73, with higher levels detected on Ig isotype-switched B cells. The few thymocytes which have detectable surface CD73 are found within CD4-CD8-and the CD4+CD8- subpopulations. In peripheral lymphoid organs, large proportions of the CD4+ and CD8+ T lymphocytes express CD73. Significant variation in the frequencies of peripheral CD73+ T cells have been observed amongst inbred mouse strains. For example, C57BL/6 mice reportedly have higher frequencies of peripheral CD73+ T cells when compared with BALB/c mice. In the thymus and peripheral lymphoid organs, CD73 is found on endothelial and stromal cells. CD73 has also been detected on bone marrow and thymic epithelial cell lines, kidney glomeruli and proximal-tubule epithelial cells, liver endothelial cells and hepatocytes, mesenchymal cells, and fibroblasts including cancer-associated fibroblasts (CAFs) found in tumors. CD73 has enzymatic and signal transduction activities. It catalyzes the dephosphorylation of extracellular nucleoside 5' monophosphates to nucleosides. CD73 acts on adenosine monophosphate (AMP) to generate and regulate the concentration of extracellular adenosine. Adenosine can bind to adenosine receptors expressed on cells in many tissues and regulate physiological responses including anti-inflammatory or immunosuppressive responses. Regulatory T cells (Treg) can generate immunosuppressive adenosine by their expression and activity of the CD39 and CD73 ectoenzymes.
Development References (6)
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Barron L, Dooms H, Hoyer KK, et al. Cutting edge: mechanisms of IL-2-dependent maintenance of functional regulatory T cells.. J Immunol. 2010; 185(11):6426-30. (Clone-specific: Flow cytometry). View Reference
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Deaglio S, Dwyer KM, Gao W, et al. Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory T cells mediates immune suppression.. J Exp Med. 2007; 204(6):1257-65. (Biology). View Reference
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Le Texier L, Lineburg KE, Cao B, et al. Autophagy-dependent regulatory T cells are critical for the control of graft-versus-host disease. JCI Insight. 2016; 1(15):e86850. (Clone-specific: Flow cytometry). View Reference
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Resta R, Yamashita Y, Thompson LF. Ecto-enzyme and signaling functions of lymphocyte CD73. Immunol Rev. 1998; 161:95-109. (Biology). View Reference
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Yamashita Y, Hooker SW, Jiang H, et al. CD73 expression and fyn-dependent signaling on murine lymphocytes. Eur J Immunol. 1998; 28(10):2981-2990. (Immunogen: Flow cytometry). View Reference
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Yu M, Guo G, Huang L, et al. CD73 on cancer-associated fibroblasts enhanced by the A2B-mediated feedforward circuit enforces an immune checkpoint.. Nat Commun. 2020; 11(1):515. (Clone-specific: Flow cytometry). View Reference
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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.