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Flow cytometric analysis of CD279 (PD-1) expression on resting and activated mouse splenic leucocytes. Splenic leucocytes from a C57BL/6 mouse were cultured without (Left Plot) or with (Right Plot) plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057) for 3 days. The resting (Left Plot) or activated (Right Plot) splenocytes were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with either BD Horizon™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984; dashed line histograms) or BD Horizon APC-R700 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 566876; solid line histograms) at 1 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histograms showing CD276 (PD-1) expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ and FlowJo® software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ APC-R700 Rat Anti-Mouse CD279 (PD-1)
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准备和存储
商品通知
- 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.
- 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.
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
配套商品
The RMP1-30 monoclonal antibody specifically recognizes CD279 which is also known as PD-1 (programmed death-1). CD279 (PD-1) is a ~55 kDa type I transmembrane glycoprotein that is encoded by Pdcd1 which belongs to the CD28/CTLA-4 family within the Ig superfamily. CD279 (PD-1) is comprised of an extracellular region with an IgV-like domain and an intracellular region with an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM) that are associated with inhibitory signaling functions. CD279 (PD-1) is transiently expressed on CD4-CD8- thymocytes and developing B lymphocytes at the pro-B-cell stage. It is also expressed on activated myeloid cells, B cells, and T cells including exhausted T cells found in mice during chronic viral infections or cancer. This co-inhibitory receptor reportedly functions in negative regulation of immune responses and thus helps guard against autoimmunity and preserves peripheral tolerance. CD273 (also known as PD-L2 or B7-DC) and CD274 (PD-L1 or B7-H1) are members of the B7 family within the Ig superfamily that serve as ligands for CD279 (PD-1). The RMP1-30 antibody reportedly does not block the binding of CD279 (PD-1) to these ligands.
This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).
研发参考 (7)
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Agata Y, Kawasaki A, Nishimura H, et al. Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int Immunol. 1996 May; 8(5):765-772. (Biology). 查看参考
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Gupta PK, Godec J, Wolski D, et al. CD39 Expression Identifies Terminally Exhausted CD8+ T Cells. PLoS Pathogens. 2015; 11(10):e1005177. (Clone-specific: Flow cytometry). 查看参考
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Kasagi S, Kawano S, Okazaki T, et al. Anti-programmed cell death 1 antibody reduces CD4+PD-1+ T cells and relieves the lupus-like nephritis of NZB/W F1 mice. J Immunol. 2010; 184(5):2337-2347. (Clone-specific: Flow cytometry). 查看参考
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Liu X, Gibbons RM, Harrington SM, et al. Endogenous tumor-reactive CD8(+) T cells are differentiated effector cells expressing high levels of CD11a and PD-1 but are unable to control tumor growth. Oncoimmunology. 2013; 2(6):e23972. (Clone-specific: Flow cytometry). 查看参考
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Matsumoto K, Inoue H, Nakano T, et al. B7-DC regulates asthmatic response by an IFN-gamma-dependent mechanism. J Immunol. 2004; 172(4):2530-2541. (Immunogen: Flow cytometry, Functional assay). 查看参考
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Nishimura H, Agata Y, Kawasaki A, et al. Developmentally regulated expression of the PD-1 protein on the surface of double-negative (CD4-CD8-) thymocytes. Int Immunol. 1996 May; 8(5):773-780. (Biology). 查看参考
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Tsushima F, Iwai H, Otsuki N, et al. Preferential contribution of B7-H1 to programmed death-1-mediated regulation of hapten-specific allergic inflammatory responses. Eur J Immunol. 2003; 33(10):2773-2782. (Biology). 查看参考
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