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  3. Fluorescent Dye
  4. DAPGreen

DAPGreen 是一种可穿透细胞的自噬荧光探针。DAPGreen 可与 PI3P 特异性结合，能标记经典自噬、替代自噬、基础自噬、线粒体自噬以及内质网自噬过程中的自噬结构，作为成像试剂可对早期自噬体和晚期自溶酶体进行染色，产生类似膜结构的“环状”信号。DAPGreen 不会影响自噬活性，在浓度高达 1.0 μM 时仍无细胞毒性，无需基因编辑即可实现活细胞成像 (Ex/Em = 453/550 nm)。

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DAPGreen

DAPGreen Chemical Structure

1.  客户无需承担相应的运输费用。

2.  同一机构（单位）同一产品试用装仅限申领一次，同一机构（单位）一年内

     可免费申领三个不同产品的试用装。

3.  试用装只面向终端客户。

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Customer Review

DAPGreen 相关抗体

Top Publications Citing Use of Products

  • 生物活性

  • 纯度 & 产品资料

  • 参考文献

生物活性

DAPGreen is a cell-permeable autophagy fluorescent probe. DAPGreen specifically binds to PI3P, and can label autophagic structures during canonical autophagy, alternative autophagy, basal autophagy, mitophagy and reticulophagy. As an imaging reagent, it stains early autophagosomes and late autolysosomes, producing a membrane-like "ring-shaped" signal. DAPGreen does not affect autophagic activity, shows no cytotoxicity even at concentrations up to 1.0 μM, and enables live-cell imaging without the need for gene editing (Ex/Em = 453/550 nm)[1][2][3].

体外研究
(In Vitro)

指南 (以下是我们推荐的方案，本方案仅提供指南，应根据您的具体需要进行修改)。
1. 染料制备
1.1 制备储存液
用无水 DMSO 配制 100 μM 的储备液。
注：储存液建议分装后于 -20 ℃ 或 -80 ℃ 避光保存。
1.2 工作液的配制
用预热好的无血清细胞培养基稀释储存液，配制成 100-500 nM 的工作液。
注：请根据实际情况调整工作液浓度，且现用现配。
2. 细胞染色
2.1 移除细胞培养基，用无血清培养基洗涤细胞两次，每次 5-10 分钟。
2.2 加入 100-500 nM 的工作液，37°C，孵育 30 分钟。
2.3 弃去上清，用无血清培养基洗涤细胞两次，每次 5-10 分钟。
2.4 将细胞置于无血清培养基中饥饿培养 2 小时至过夜，以诱导自噬。并设置适当的对照组。
2.5. 使用荧光显微镜或流式细胞仪观察荧光信号。


DAPGreen (0.25 μM; 30 min prestain, 5 h starvation) 可标记饥饿处理的 MEF 细胞中的经典自噬结构；在 Atg5 敲除 MEF 细胞 (可形成早期吞噬泡) 中可检测到信号，而在 Atg9 敲除和 Ulk1/2 双敲除 MEF 细胞 (无法形成早期吞噬泡) 中信号极弱[1]。
DAPGreen (5.0 μM) 在无细胞缓冲液体系中，于 pH 4.0-8.0 范围内表现出不依赖 pH 的荧光特性[2]。
DAPGreen (0.1 μM; 30 min preincubation, 4 h autophagy induction) 可在营养饥饿的 HeLa 细胞中对早期自噬体和晚期自溶酶体进行染色，在 0.1 μM 浓度下自噬诱导 4 h 的过程中其荧光强度逐渐增强[2]。
DAPGreen (0.5 μM; 30 min preincubation, 4 h rapamycin treatment) 在 Rapamycin 处理的 HeLa 细胞中以 0.5 μM 浓度使用时，会与自噬体标记物 tagRFP-LC3 共定位[2]。
DAPGreen (0.1-1.0 μM; 30 min incubation, 24 h post-incubation) 在 0.1 μM、0.5 μM 和 1.0 μM 浓度下对 HeLa 细胞无细胞毒性[2]。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

DAPGreen 相关抗体:

Immunofluorescence[1]

Cell Line: HeLa cells
Concentration: 0.1 μM (DAPGreen preincubation); 0.1 μM (BafA1)
Incubation Time: 30 min (DAPGreen preincubation); 3 h (starvation with/without BafA1)
Result: Showed weak fluorescence in untreated control cells.
Increased significantly in fluorescent area per cell in starved cells.
Remained present after BafA1 treatment, with a larger fluorescent area than control cells but smaller than starved-only cells.

Immunofluorescence[1]

Cell Line: WT MEFs transfected with mCherry-WIPI2
Concentration: 0.25 μM (DAPGreen); 0.5 μM (rapamycin)
Incubation Time: 30 min (DAPGreen preincubation); 3 h (rapamycin treatment)
Result: Labeled all WIPI2 puncta (phagophore structures), with a colocalization rate of 90% in rapamycin-treated cells.

Immunofluorescence[1]

Cell Line: WT MEFs transfected with TagRFP-LC3
Concentration: 0.25 μM (DAPGreen); 0.5 μM (rapamycin)
Incubation Time: 30 min (DAPGreen preincubation); 3 h (rapamycin treatment)
Result: Almost completely labeled TagRFP-LC3 puncta, which represent autophagic structures from phagophores through autolysosomes, with a colocalization rate of 95%.

Immunofluorescence[1]

Cell Line: WT, Atg5 KO, Atg9 KO, and Ulk1/2 DKO MEFs
Concentration: 0.25 μM (DAPGreen)
Incubation Time: 30 min (DAPGreen prestain); 5 h (starvation)
Result: Showed the highest mean total fluorescent area per cell (220 μm2) in WT MEFs.
Showed a reduced but detectable fluorescent area (50 μm2) in Atg5 KO MEFs.
Showed further reduced areas (20 μm2 and 10 μm2, respectively) in Atg9 KO and Ulk1/2 DKO MEFs.
All KO groups showed statistically significant reductions compared to WT, and Atg5 KO MEFs had a significantly larger area than Atg9 KO and Ulk1/2 DKO MEFs.

Immunofluorescence[1]

Cell Line: WT MEFs
Concentration: 0.25 μM (DAPGreen); 0.1 μM (DAPRed); 0.5 μM (rapamycin)
Incubation Time: 30 min (DAPGreen and DAPRed prestain); 2 h (rapamycin treatment for static imaging); 47 s (time-lapse imaging)
Result: Colocalized with all DAPRed puncta, while a small subset of DAPGreen-only puncta (early phagophores) was observed.
Showed DAPRed signals emerging and expanding over pre-existing DAPGreen puncta within 47 s via time-lapse imaging.

Cell Viability Assay[2]

Cell Line: HeLa cells
Concentration: 0.1-1.0 μM
Incubation Time: 30 min (DAPGreen incubation); 24 h (incubation without DAPGreen)
Result: Showed no cytotoxicity in HeLa cells at concentrations up to 1.0 μM, as measured by CCK-8 absorbance.
体内研究
(In Vivo)

DAPGreen (0.1 mM; 水中浸泡；30 min) 由于受到生物自发荧光的干扰，无法用于检测活体斑马鱼中的自噬结构[1]。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

459.02

Formula

C25H35ClN4O2
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.
纯度 & 产品资料
参考文献

DAPGreen 相关分类

  • 摩尔计算器

  • 稀释计算器

The molarity calculator equation

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

质量   浓度   体积   分子量 *
= × ×

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

浓度 (start) × 体积 (start) = 浓度 (final) × 体积 (final)
× = ×
C1   V1   C2   V2
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

DAPGreenFluorescent DyeAtg5 KO MEFsHeLa cellsmouse embryosphosphatidylinositol 3-phosphatePI3PautophagosomesautolysosomesphagophoresWT MEFsautophagyInhibitorinhibitorinhibit

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