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BD Horizon™ Violet Proliferation Dye 450
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BD Horizon™ Violet Proliferation Dye 450

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品牌: BD Pharmingen
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实验应用:
Intracellular staining (flow cytometry) (Routinely Tested)
产品介绍
产品介绍
产品信息
简单描述
BD Horizon™ Violet Proliferation Dye 450 (VPD450) is a violet laser excitable dye that can be used for flow cytometric monitoring of cell divisions completed by single cells. The nonfluorescent VPD450 dye passively diffuses across cell membranes and is cleaved by esterase activity within viable cells. The cleaved dye becomes highly fluorescent and covalently binds to protein amine groups within the cells via its succinimidyl ester group. Nonviable cells remain nonfluorescent. As viable cells divide, the VPD450 dye is distributed uniformly between daughter cells; each daughter cell retains approximately half of the VPD450 fluorescence intensity of its parent cell. The covalently bound dye is well retained within the cells allowing for applications involving live or aldehyde-fixed and permeabilized cells.          The violet laser excitable dye VPD450 emits maximally at 450 nm with minimal fluorescent spillover into the FITC channel. This allows for use with green fluorescent protein (GFP)-tagged cells and in other multicolor assays requiring measurements in the FITC channel, such as intracellular cytokine assays.
商品描述
BD Horizon™ Violet Proliferation Dye 450 (VPD450) is a violet laser excitable dye that can be used for flow cytometric monitoring of cell divisions completed by single cells. The nonfluorescent VPD450 dye passively diffuses across cell membranes and is cleaved by esterase activity within viable cells. The cleaved dye becomes highly fluorescent and covalently binds to protein amine groups within the cells via its succinimidyl ester group. Nonviable cells remain nonfluorescent. As viable cells divide, the VPD450 dye is distributed uniformly between daughter cells; each daughter cell retains approximately half of the VPD450 fluorescence intensity of its parent cell. The covalently bound dye is well retained within the cells allowing for applications involving live or aldehyde-fixed and permeabilized cells. The violet laser excitable dye VPD450 emits maximally at 450 nm with minimal fluorescent spillover into the FITC channel. This allows for use with green fluorescent protein (GFP)-tagged cells and in other multicolor assays requiring measurements in the FITC channel, such as intracellular cytokine assays.
克隆号
(RUO)
应用
实验应用
Intracellular staining (flow cytometry) (Routinely Tested)
目标/特异性
Violet Cell Proliferation Dyes
文献
文献
研发参考(3) 1. Lyons AB, Doherty KV. Flow cytometric analysis of cell division by dye dilution. Curr Protoc Cytom. 2004; 9(Unit 9.11):1-9. (Biology). 2. Lyons AB. Analysing cell division in vivo and in vitro using flow cytometric measurement of CFSE dye dilution. J Immunol Methods. 2000; 243(1-2):147-154. (Biology). 3. Parish CR. Fluorescent dyes for lymphocyte migration and proliferation studies. Immunol Cell Biol. 1999; 77(6):499-508. (Methodology: Flow cytometry).

参考图片

Figure 2. Staining of surface and intracellular phenotypes on activated mouse splenocytes. A single-cell suspension of mouse splenocytes was treated to remove red blood cells and then labeled with VPD450 (1 μM) for 10 minutes. The cells were washed twice and then cultured with Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in complete tissue culture medium. After 2 days, the cells were harvested, washed, and restimulated with Phorbol 12-Myristate 13-Acetate and Ionomycin in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 4 hrs. The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat No. 554715) followed by staining with PerCP-Cy™5.5 Anti-Mouse CD4 (Cat. No. 550954) and FITC Anti-Mouse IL-2 (Cat. No. 554427) antibodies. Two-color flow cytometric dot plots showing the correlated expression patterns of VPD450 versus CD4 (Left Panel) or IL-2 (Right Panel) were derived from gated events with the forward and side light-scatter characteristics of intact splenic lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Figure 1. Flow cytometric analysis of proliferative responses by human peripheral blood lymphocytes. A single-cell suspension of human peripheral blood mononuclear cells was labeled with VPD450 (1 μM) for 10 minutes. The cells were washed twice and then cultured with phytohemagglutinin for four days. The histogram shows VPD450 fluorescence peaks of gated events with the forward and side light-scatter characteristics of viable lymphocytes (successive generations of divided cells). Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Figure 2. Staining of surface and intracellular phenotypes on activated mouse splenocytes. A single-cell suspension of mouse splenocytes was treated to remove red blood cells and then labeled with VPD450  (1 μM) for 10 minutes. The cells were washed twice and then cultured with Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057) and Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294) antibodies in complete tissue culture medium. After 2 days, the cells were harvested, washed, and restimulated with Phorbol 12-Myristate 13-Acetate and Ionomycin in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 4 hrs. The cells were then fixed and permeabilized using a BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat No. 554715) followed by staining with PerCP-Cy™5.5 Anti-Mouse CD4 (Cat. No. 550954) and FITC Anti-Mouse IL-2 (Cat. No. 554427) antibodies. Two-color flow cytometric dot plots showing the correlated expression patterns of VPD450 versus CD4 (Left Panel) or IL-2 (Right Panel) were derived from gated events with the forward and side light-scatter characteristics of intact splenic lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

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货号:
562158
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