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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. 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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- CF™ is a trademark of Biotium, Inc.
- 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 BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
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.
This antibody is conjugated to BD Horizon™ BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
Development References (6)
-
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
-
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
-
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
-
Resta R, Yamashita Y, Thompson LF. Ecto-enzyme and signaling functions of lymphocyte CD73. Immunol Rev. 1998; 161:95-109. (Biology). View Reference
-
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
-
Yu M, Guo G, Huang L, et al. CD73 on cancer-associated fibroblasts enhanced by the A(2B)-mediated feedforward circuit enforces an immune checkpoint. Nature Commun. 2020; 11(1):515. (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.
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.
