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Two-color flow cytometric analysis of CD27 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either BD Horizon™ BV737 Hamster IgG1, κ Isotype Control (Cat. No. 564621; Left Plot) or BD Horizon BV737 Hamster Anti-Mouse CD27 antibody (Cat. No. 565307; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of CD27 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BUV737 Hamster Anti-Mouse CD27
Regulatory Status Legend
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
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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The LG.3A10 monoclonal antibody specifically binds to CD27, a lymphocyte-restricted member of the Tumor Necrosis Factor Receptor family which binds to CD70. The CD27 molecule is a 45-kDa transmembrane glycoprotein which is constitutively expressed by lymphocytes of the T lineage: virtually all thymocytes and over 90% of peripheral T cells bearing both αβ and γδ T-cell receptors. CD27 cooperates with the pre-TCR in mediating thymocyte differentiation and expansion. In addition, one to ten percent of mature peripheral B cells express CD27, and CD27's role in the differentiation of human plasma cells has been studied. Mouse NK cells, freshly isolated and IL-2-activated, also express CD27. In the bone marrow, CD27 is found on a progenitor population which provides short-term hematopoietic reconstitution. Cells of the myeloid lineage do not express CD27. Cross-linked LG.3A10 mAb has been reported to amplify the proliferative response of purified T lymphocytes to suboptimal mitogenic stimulation1 and to enhance NK-cell proliferation and IFN-γ production. In contrast, non-cross-linked LG.3A10 mAb inhibits CD3-induced pre-T cell development by interfering with the receptor-ligand interaction. This hamster mAb to a mouse leukocyte antigen has been observed to cross-react with a similar population of rat leukocytes.
Development References (6)
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Agematsu K, Hokibara S, Nagumo H, Shinozaki K, Yamada S, Komiyama A. Plasma cell generation from B-lymphocytes via CD27/CD70 interaction. Leuk Lymphoma. 1999; 35(3-4):219-225. (Biology). View Reference
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Gravestein LA, Blom B, Nolten LA, et al. Cloning and expression of murine CD27: comparison with 4-1BB, another lymphocyte-specific member of the nerve growth factor receptor family. Eur J Immunol. 1993; 23(4):943-950. (Biology). View Reference
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Gravestein LA, Nieland JD, Kruisbeek AM, Borst J. Novel mAbs reveal potent co-stimulatory activity of murine CD27. Int Immunol. 1995; 7(4):551-557. (Immunogen: (Co)-stimulation, Flow cytometry, Functional assay, Immunofluorescence, Immunoprecipitation). View Reference
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Gravestein LA, van Ewijk W, Ossendorp F, Borst J. CD27 cooperates with the pre-T cell receptor in the regulation of murine T cell development. J Exp Med. 1996; 184(2):675-685. (Clone-specific: Flow cytometry, Immunofluorescence, Inhibition). View Reference
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Takeda K, Oshima H, Hayakawa Y, et al. CD27-mediated activation of murine NK cells. J Immunol. 2000; 164(4):1741-1745. (Clone-specific: (Co)-stimulation, Enhancement, Flow cytometry). View Reference
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Wiesmann A, Phillips RL, Mojica M, et al. Expression of CD27 on murine hematopoietic stem and progenitor cells. Immunity. 2000; 12(2):193-199. (Clone-specific: Flow cytometry). View Reference
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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.