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Multiparameter flow cytometric analysis of CD27 expression on Human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or BD OptiBuild™ BV421 Mouse anti-Human CD27 antibody (Cat. No. 750853; Right Plot) at 0.25 µg/test. After staining, the cells were treated with 1× BD Pharm Lyse™ Lysing Solution (Cat. No. 555899) to remove erythrocytes. The bivariate pseudocolor density plot showing the correlated expression of CD27 (or Ig Isotype control staining) versus side light-scatter signals was derived from gated events with the light scattering characteristics for intact leucocyte populations. Flow cytometric analysis was performed using a BD Celesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD OptiBuild™ BV421 Mouse Anti-Human CD27
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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.
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/) 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).
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- 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.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The O323 monoclonal antibody specifically recognizes CD27 which is also known as Tumor necrosis factor receptor superfamily member 7 (TNFRSF7), T14, Tp55, or S152. CD27 exists as a ~110-120 kDa disulfide-linked homodimer comprised of two single-pass type I transmembrane glycoproteins that are encoded by CD27 (CD27 molecule). CD27 is expressed on medullary thymocytes and T cells, with higher expression on activated T cells, and subsets of mature B cells and natural killer (NK) cells. A soluble 28-32 kDa form of CD27 is produced by lymphocytes upon cellular activation. Binding of the CD27 antigen, expressed on T cells, to its ligand, CD70 (CD27L), provides a costimulatory signal, leading to T cell proliferation, production of cytotoxic T cells, and enhanced production of cytokines. Binding of CD70 to CD27 expressed on B cells leads to B cell proliferation and the generation of plasma cells and immunoglobulin production. The CD27 antigen becomes hyperphosphorylated on serine residues upon activation of T cells. Signaling through the CD27 antigen activates NFκB and stress activated protein kinase (SAPK)/c Jun N terminal kinase (JNK).
Development References (5)
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Björkström NK, Béziat V, Cichocki F, et al. CD8 T cells express randomly selected KIRs with distinct specificities compared with NK cells.. Blood. 2012; 120(17):3455-65. (Clone-specific: Flow cytometry). View Reference
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Borst J, Hendriks J, Xiao Y. CD27 and CD70 in T cell and B cell activation.. Curr Opin Immunol. 2005; 17(3):275-81. (Biology). View Reference
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Klein U, Rajewsky K, Küppers R. Human immunoglobulin (Ig)M+IgD+ peripheral blood B cells expressing the CD27 cell surface antigen carry somatically mutated variable region genes: CD27 as a general marker for somatically mutated (memory) B cells.. J Exp Med. 1998; 188(9):1679-89. (Biology). View Reference
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Reiter C. T9. Cluster report: CD27. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:350.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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