Fixable Viability Stain 450

品牌： BD Pharmingen

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实验应用：

Intracellular staining (flow cytometry), Flow cytometry (Tested During Development)

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简单描述

BD Horizon™ Fixable Viability Stain 450 (FVS450) is useful to discriminate viable from non-viable mammalian cells in multicolor flow cytometric applications. This violet fluorescent stain contains a dye that reacts with and covalently binds to cell surface and intracellular amines. Permeable plasma cell membranes, such as those present in necrotic cells, allow for the intracellular diffusion of the violet dye and covalent binding to higher overall concentrations of amines than in non-permeable live cells. Therefore, necrotic cells present in a typical in vitro assay label with higher levels of dye increasing their fluorescence intensity 10-20 fold over that of viable cells. The labeled cells can be fixed with formaldehyde for downstream decontamination, freezing and/or permeablization and subsequent intracellular staining while maintaining stable FVS450 fluorescence. The BD Horizon™ Fixable Viability Stain 450 is excited by the Violet laser (with an excitation maximum of 406 nm) and has a fluorescence emission maximum at 450 nm.

商品描述

应用

实验应用

Intracellular staining (flow cytometry), Flow cytometry (Tested During Development)

文献

研发参考(4) 1. Abrams B, Diwu Z, Guryev O, et al. 3-Carboxy-6-chloro-7-hydroxycoumarin: a highly fluorescent, water-soluble violet-excitable dye for cell analysis. Anal Biochem. 2009; 386(2):262-269. (Methodology). 2. Burmeister Y, Lischke T, Dahler AC, et al. ICOS controls the pool size of effector-memory and regulatory T cells. J Immunol. 2008; 180(2):774-782. (Methodology). 3. Charles ED, Green RM, Marukian S, et al. Clonal expansion of immunoglobulin M+CD27+ B cells in HCV-associated mixed cryoglobulinemia. Blood. 2008; 111(3):1344-1356. (Methodology). 4. Perfetto SP, Chattopadhyay PK, Lamoreaux L, et al. Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry. J Immunol Methods. 2006; 313(1–2):199-208. (Methodology).

参考图片

Multicolor flow cytometric analysis of phosphorylated STAT3 expression by “viable” activated human peripheral blood mononuclear cells (PBMC). PBMC were cultured for 48 hours in complete tissue culture medium and then frozen and stored (-80°C) for ten days. The cells were thawed and treated with recombinant human IL-6 (100 ng/ml; Cat. No. 550071) for 15 minutes with BD Horizon™ Fixable Viability Stain 450 (Cat. No. 562247) added for the last 7 minutes of activation. Cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) according to the standard Phosflow protocol. Cells were stained with PE Mouse Anti-Human CD3 (Cat. No. 555333), PerCP-Cy™5.5 Mouse Anti-Human CD4 (Cat. No. 552838) and BD Phosflow™ Alexa Fluor® 647 Mouse Anti-Stat3 (pY705) (Cat. No. 557815) antibodies. The dual parameter flow cytometric dot plot (Left Panel) shows the incorporated levels of FVS450 versus side scattered light signals expressed by the PBMC. Flow cytometric histograms show the levels of Stat3 (pY705) expressed by live cell-discriminated (ie, gated events with low level FVS450 incorporation; Middle Panel) and total events (including cells with both low and high levels of FVS450; Right Panel). The CD4+CD3+ T lymphocytes were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System. FVS450 was also tested in mouse (data not shown).

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562247

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