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Multiparameter flow cytometric analysis of CD27 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD27 antibody (Cat. No. 567025/567026; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD27 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Human CD27
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.