Flow cytometry, Intracellular staining (flow cytometry) (Tested During Development)
产品介绍
产品介绍
产品信息
简单描述
BD Horizon™ Fixable Viability Stain 520 (FVS520) is useful for discrimination of viable from non-viable mammalian cells in multicolor flow cytometric applications. This dye 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 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 permeabilization and subsequent intracellular staining while maintaining stable viability stain fluorescence.
BD Horizon™ Fixable Viability Stain 520 is excited by the Blue laser (with an excitation maximum of 498 nm) and has a fluorescence emission maximum of 521 nm.
商品描述
BD Horizon™ Fixable Viability Stain 520 (FVS520) is useful for discrimination of viable from non-viable mammalian cells in multicolor flow cytometric applications. This dye 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 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 permeabilization and subsequent intracellular staining while maintaining stable viability stain fluorescence.
BD Horizon™ Fixable Viability Stain 520 is excited by the Blue laser (with an excitation maximum of 498 nm) and has a fluorescence emission maximum of 521 nm.
克隆号
(RUO)
应用
实验应用
Flow cytometry, Intracellular staining (flow cytometry) (Tested During Development)
目标/特异性
Live/Dead Discriminator Dyes
文献
文献
研发参考(5)
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).
5. Perfetto SP, Chattopadhyay PK, Lamoreaux L, et al. Amine-reactive dyes for dead cell discrimination in fixed samples. Curr Protoc Cytom. 9(9.34)(Methodology).
参考图片
Flow cytometric analysis of human Jurkat cells stained with BD Horizon™ Fixable Viability Stain 520. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were treated with 0.025% DMSO (Left Panel) or 5 μM camptothecin (Right Panel) for 16 hours and then stained with BD Horizon™ Fixable Viability Stain 520 (Cat. No. 564407) in serum-free buffer. The cells were then either left unfixed (solid line histograms) or fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized in BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) (dashed line histograms). Histograms were derived from gated events with the light scattering characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Please note that FVS520 is also compatible with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725).
Flow cytometric analysis of human Jurkat cells stained with BD Horizon™ Fixable Viability Stain 520. Cells from the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line were treated with 0.025% DMSO (Left Panel) or 5 μM camptothecin (Right Panel) for 16 hours and then stained with BD Horizon™ Fixable Viability Stain 520 (Cat. No. 564407) in serum-free buffer. The cells were then either left unfixed (solid line histograms) or fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized in BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) (dashed line histograms). Histograms were derived from gated events with the light scattering characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Please note that FVS520 is also compatible with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). Fixable Viability Stain 520 has been tested on mouse (data not shown).
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