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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 PE-Cy™7 Hamster IgG1, κ Isotype Control (Cat. No. 552811; Left Panel) or PE-Cy™7 Hamster Anti-Mouse CD27 antibody (Cat. No. 563604; Right Panel). Two-color flow cytometric dot plots showing the correlated expression patterns 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 Pharmingen™ PE-Cy™7 Hamster Anti-Mouse 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
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
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- 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.
- 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.
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- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
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- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. 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 PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- 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.
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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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.