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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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 OX-22 antibody reacts with CD45RC on pre-B lymphocytes, B cells, CD8+ T suppressor/cytotoxic cells, and a subset of CD4+ T helper (Th) lymphocytes. It weakly reacts with thymocytes. CD45RC is a high-molecular-weight isoform of CD45 (Leukocyte Common Antigen); its level of expression distinguishes subpopulations of CD4+ T cells with Th1-like and Th2-like effector functions. Levels of expression of CD45RC have also been reported to distinguish resting from activated T cells at various stages of maturation. CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family: Its intracellular (COOH-terminal) region contains two PTP catalytic domains, and the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), plus differing levels of glycosylation. The CD45 isoforms detected in the rat are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction.
Development References (14)
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Arthur RP, Mason D. T cells that help B cell responses to soluble antigen are distinguishable from those producing interleukin 2 on mitogenic or allogeneic stimulation. J Exp Med. 1986; 163(4):774-786. (Biology). View Reference
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Bunce C, Bell EB. CD45RC isoforms define two types of CD4 memory T cells, one of which depends on persisting antigen. J Exp Med. 1997; 185(4):767-776. (Biology). View Reference
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Fowell D, McKnight AJ, Powrie F, Dyke R, Mason D. Subsets of CD4+ T cells and their roles in the induction and prevention of autoimmunity. Immunol Rev. 1991; 123:37-64. (Biology). View Reference
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Groen H, Klatter FA, van Petersen AS, Pater JM, Nieuwenhuis P, Kampinga J. Composition of rat CD4+ resting memory T-cell pool is influenced by major histocompatibility complex. Transplant Proc. 1993; 25(5):2782-2783. (Biology). View Reference
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Hargreaves M, Bell EB. Identical expression of CD45R isoforms by CD45RC+ 'revertant' memory and CD45RC+ naive CD4 T cells. Immunology. 1997; 91(3):323-330. (Biology). View Reference
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Johnson P, Maiti A, Ng DHW. CD45: A family of leukocyte-specific cell surface glycoproteins. In: Herzenberg LA, Weir DM, Herzenberg LA, Blackwell C , ed. Weir's Handbook of Experimental Immunology, Vol 2. Cambridge: Blackwell Science; 1997:62.1-62.16.
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Kampinga J, Groen H, Klatter F, et al. Post-thymic T cell development in rats: an update. Biochem Soc Trans. 1992; 20(1):191-197. (Biology). View Reference
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Mason DW, Arthur RP, Dallman MJ, Green JR, Spickett GP, Thomas ML. Functions of rat T-lymphocyte subsets isolated by means of monoclonal antibodies. Immunol Rev. 1983; 74:57-82. (Clone-specific). View Reference
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Papp I, Wieder KJ, Sablinski T, et al. Evidence for functional heterogeneity of rat CD4+ T cells in vivo. Differential expression of IL-2 and IL-4 mRNA in recipients of cardiac allografts. J Immunol. 1992; 148(5):1308-1314. (Biology). View Reference
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Sarawar SR, Sparshott SM, Sutton P, Yang CP, Hutchinson IV, Bell EB. Rapid re-expression of CD45RC on rat CD4 T cells in vitro correlates with a change in function. Eur J Immunol. 1993; 23(1):103-109. (Biology). View Reference
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Sparshott SM, Bell EB. Membrane CD45R isoform exchange on CD4 T cells is rapid, frequent and dynamic in vivo. Eur J Immunol. 1994; 24(11):2573-2578. (Biology). View Reference
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Spickett GP, Brandon MR, Mason DW, Williams AF, Woollett GR. MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte-common antigen. J Exp Med. 1983; 158(3):795-810. (Immunogen). View Reference
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Woollett GR, Barclay AN, et al. Molecular and antigenic heterogeneity of the rat leukocyte common antigen from thymocytes and T and B lymphocytes. . Eur J Immunol. 1985; 15:168-173. (Clone-specific).
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Yang CP, Bell EB. Functional maturation of recent thymic emigrants in the periphery: development of alloreactivity correlates with the cyclic expression of CD45RC isoforms. Eur J Immunol. 1992; 22(9):2261-2269. (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.