计算溶液所需的质量、体积或浓度。
aladdin 488：Ex/Em = 500/530 nm (with DNA)
有货
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
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避光,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
aladdin™ 488 Caspase-3 Substrate 基于caspase-3/7 活力检测细胞凋亡提供了有效的工具，适用于荧光显微术和流式细胞术。 相比其他基于（FLICA）分析的 caspase 的荧光底物或荧光抑制剂，aladdin™ 488 Caspase-3 Substrate 检测 caspase-3/7 活性的同时不会抑制完整细胞的凋亡过程。 Substrate 由耦合caspase-3/7 DEVD识别序列的荧光 DNA染料组成。Substrate 最初无荧光，穿过细胞膜进入细胞质。在凋亡细胞中，caspase-3/7剪切Substrate，释放高亲和性的DNA染色，这种染料迁移到细胞核标记DNA并发出 明亮的绿色荧光。因此，aladdin™ 488 Caspase-3 Substrate 是双功能的，既可以检测caspase-3/7活性，又可视化细胞核在细胞凋亡进行中的形态学变化。aladdin™ 488染色可以甲醛固定并兼容后续免疫染色实验。 aladdin™ 488 Caspase-3 Substrate 提供 DMSO 和 PBS 两种溶解形式。PBS 形式可用于对 DMSO 毒性敏感的细胞，在对 DMSO 不敏感的细胞类型中，添加 DMSO 溶解形式可增强、aladdin™ 488 的孵育染色效果。
产品参数：
aladdin 488：Ex/Em = 500/530 nm (with DNA)
注意事项：
1. 细胞可以用终浓度为1 μM的Hoechst 33342染料共同染色，使细胞核产生蓝色荧光染色（激发/发射：346/460 nm）。
2. aladdin 488 染色可以被甲醛固定，但与甲醇固定不兼容。
3. 甲醛固定的 aladdin 488 染色细胞可以用 0.1% Triton X-100 处理后进行后续染色，但这样处理后的染色的亮度可能会减弱。
应用范围：
Caspase-3底物、酶底物
使用方法：
1. 实验优化
下 面 提 供 的 实 验 步 骤 根 据 终 点 法 检 测 制 度 。 aladdin 488 Substrate也可以进行细胞长时间孵育研究 。细胞密度、底物浓度和抑制剂浓 度可能需要优化。最佳底物浓度可能在1-10 μM之间。细胞 可以在培养基、PBS或其他您所选择的缓冲液中孵育底物。 对于贴壁细胞，我们建议更换新鲜含有底物的培养基，以防 背景的不均一性。底物孵育后换液或洗涤细胞的操作是自由 选择的。当在DMSO不敏感的细胞类型中使用时，添加DMSO可能会加强 aladdin 488染色效果。
2. 对照 我们建议您设定以下对照：
A. 阴性对照：不诱导凋亡的细胞
B. 阳性对照：诱导凋亡的细胞
3. 流式细胞术
（1）选择合适的方法诱导细胞凋亡，未经处理的细胞样本 作为对照。
（2）贴壁细胞，进行SuperView™ 488 Caspase-3实验前先用 胰蛋白酶或其他方法消化细胞。
（3）用培养基或缓冲液重悬细胞，使细胞密度为106 /mL。
（4）吸取0.2 mL细胞悬液至流式细胞试管。
（5）往200 μL细胞悬液中加入1 μL 1 mM的Substrate 并立 即混匀使底物浓度为5 μM。不同细胞的最佳底物浓度可能 不同，需分析优化。
（6）室温避光孵育细胞15-30 min。
（7）加入300 μL培养基或PBS，流式细胞仪分析。检测绿色 荧光的通道（激发/发射：485/515 nm）。
4. 荧光显微镜
（1）选择合适的方法诱导细胞凋亡，未经处理的细胞样本作为对照。
（2）用含有5 μM Substrate 的新鲜培养基或PBS对细胞进行换液（见试验优化）。
（3）室温下孵育细胞30 min或更长时间。
（4）细胞可以在含有Substrate的培养基中直接观察。对于终点分析法，PBS清洗细胞，荧光显微镜观察细胞，使用观察 绿色荧光的滤片（激发/发射：485/515 nm）。
5. 荧光酶标仪
（1）贴壁细胞在黑色96孔板中进行培养；悬浮细胞，调整 密度至106 cells/mL，0.2 mL细胞悬液分装到一孔。（2）选择合适的方法诱导细胞凋亡，未经处理的细胞样本 作为对照。注意：细胞可以在管或瓶中处理，然后转移到96 孔检测板。
（3）对于悬浮细胞，直接添加Substrate混匀。对于贴壁细胞， 用含有5 μM Substrate的新鲜培养基或PBS对细胞进行换液 （见试验优化）。
（4）室温避光孵育细胞15-30 min。
（5）对于悬浮细胞，轻轻摇晃重悬细胞。荧光酶标仪设置 激发波长488 nm和发射波长520 nm。建议贴壁细胞使用底部采集方式。贴壁细胞密度的变化可能导致不准确的读数。
常见问题
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Superview ™ 488 caspase-3 substrate provides an effective tool for detecting apoptosis based on caspase-3/7 viability, which is suitable for fluorescence microscopy and flow cytometry. Compared with other fluorescent substrates or fluorescent inhibitors of caspases based on (Flica) analysis, superview ™ 488 caspase-3 substrate does not inhibit the apoptosis process of intact cells while detecting caspase-3/7 activity. Substrate consists of a fluorescent DNA dye that couples the caspase-3/7 devd recognition sequence. Substrate is initially nonfluorescent and crosses the cell membrane into the cytoplasm. In apoptotic cells, caspase-3/7 cleaves substrate and releases high affinity DNA staining, which migrates to the nucleus to label DNA and emits bright green fluorescence. Therefore, superview ™ 488 caspase-3 substrate is bifunctional, which can not only detect caspase-3/7 activity, but also visualize the morphological changes of the nucleus during apoptosis. Superview ™ 488 staining can be formaldehyde fixed and compatible with subsequent immunostaining experiments. Superview ™ 488 caspase-3 substrate provides two dissolved forms, DMSO and PBS. The PBS form can be used for cells sensitive to DMSO toxicity, and in cell types insensitive to DMSO, the addition of DMSO dissolved form can enhance, superview ™ 488 incubation staining effect.
Product parameters：
aladdin 488：Ex/Em = 500/530 nm (with DNA)
Matters needing attention：
1. cells can be treated with a final concentration of 1 μ M Hoechst 33342 dye CO staining, resulting in blue fluorescent staining of the nucleus (excitation / emission: 346 / 460 nm). 2. superview ™ 488 staining can be fixed by formaldehyde, but is incompatible with methanol fixation.
3. formaldehyde fixed superview ™ 488 stained cells can be treated with 0.1% Triton X-100 for subsequent staining, but the brightness of the staining after such treatment may be weakened.
Scope of application：
Caspase-3 substrate, enzyme substrate
Usage:
1. Experimental optimization
The experimental steps provided below are based on the endpoint detection system. Aladdin 488 Substrate can also be used for long-term cell incubation studies. Cell density, substrate concentration, and inhibitor concentration may need to be optimized. The optimal substrate concentration may be between 1-10 μ Between M. Cells can be incubated with substrates in culture medium, PBS, or other buffer of your choice. For adherent cells, we recommend replacing them with fresh culture media containing substrates to prevent background heterogeneity. The operation of changing the medium or washing the cells after substrate incubation is freely selectable. When used in cell types that are not sensitive to DMSO, adding DMSO may enhance the staining effect of Aladdin 488.
2. We suggest that you set the following controls:
A. Negative control: cells that do not induce apoptosis
B. Positive control: Cells that induce apoptosis
3. Flow cytometry
(1) Choose appropriate methods to induce cell apoptosis, with untreated cell samples as controls.
(2) Adhering cells for SuperView ™ Before conducting the 488 Caspase-3 experiment, the cells were digested using trypsin or other methods.
(3) Resuspend cells with culture medium or buffer to achieve a cell density of 106/mL.
(4) Suck 0.2 mL of cell suspension into a flow cytometry test tube.
(5) To 200 μ Add 1 to L cell suspension μ Substrate of L 1 mM and immediately mix to achieve a substrate concentration of 5 μ M. The optimal substrate concentration for different cells may vary and requires analysis and optimization.
(6) Incubate cells at room temperature in dark for 15-30 minutes.
(7) Join 300 μ L-medium or PBS, analyzed by flow cytometry. Channel for detecting green fluorescence (excitation/emission: 485/515 nm).
4. Fluorescence microscope
(1) Choose appropriate methods to induce cell apoptosis, with untreated cell samples as controls.
(2) Using a solution containing 5 μ Fresh culture medium or PBS from M Substrate was used to replace the cells (see experimental optimization).
(3) Incubate cells at room temperature for 30 minutes or longer.
(4) Cells can be directly observed in culture media containing Substrate. For endpoint analysis, PBS is used to clean cells, fluorescence microscopy is used to observe cells, and a filter (excitation/emission: 485/515 nm) is used to observe green fluorescence.
5. Fluorescence enzyme-linked immunosorbent assay (ELISA) reader
(1) Cultivate adherent cells in black 96 well plates; Suspend cells, adjust the density to 106 cells/mL, and divide 0.2 mL of cell suspension into one well. (2) Choose appropriate methods to induce cell apoptosis, with untreated cell samples as controls. Note: Cells can be processed in tubes or bottles and then transferred to a 96 well detection plate.
(4) Incubate cells at room temperature in dark for 15-30 minutes.
(5) For suspended cells, gently shake to resuspend the cells. The fluorescence enzyme-linked immunosorbent assay instrument is set with an excitation wavelength of 488 nm and an emission wavelength of 520 nm. Suggest using bottom collection method for adherent cells. Changes in the density of adherent cells may lead to inaccurate readings.
Frequently asked question
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规格
别名
阿拉丁 488：Ex/Em = 500/530 nm (含 DNA)
规格或纯度
1 mM in DMSO
英文名称
aladdin 488：Ex/Em = 500/530 nm (with DNA)
储存条件
避光,-20°C储存
运输条件
超低温运输
名称和识别符
分子类型
小分子
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分析证书 | S598357 |
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