计算溶液所需的质量、体积或浓度。
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
细胞凋亡是指为维持内环境稳定，由基因控制的细胞自主的有序的死亡。细胞凋亡与细胞坏死不同，细细胞凋亡（Apoptosis）一般是指机体细胞在发育过程中或在某些因素作用下通过细胞内基因及其产物的调控而发生的一种程序性细胞死亡过程，细胞坏死是细胞受到强烈理化或生物因素作用引起细胞无序变化的死亡过程。细胞凋亡与坏死的区别在于特征性的形态学和生物化学变化，包括细胞膜通透性与核染色质的变化，细胞质的收缩及膜不对称的丧失。 本公司生产的Oxazole yellow/PI膜透性凋亡检测试剂盒，是一种基于Oxazole yellow和PI两种染料的双荧光法检测凋亡的试剂盒。本试剂盒适用于荧光显微镜、流式细胞仪、荧光酶标仪及其它荧光检测系统。 Oxazole yellow是一种非细胞膜穿透性的并对DNA具有高亲和力的羰花青单体绿色荧光染料，在没有与DNA结合的时候基本上没有荧光，而与DNA结合后可以发出明亮的绿色荧光。在细胞凋亡发生时，细胞膜通透性发生改变，此时Oxazole yellow可以进入细胞内与DNA结合，发出明亮的绿色荧光，因此常用于细胞凋亡的检测。需要注意的是Oxazole yellow也能染色死细胞，因此需要和对死细胞特异性荧光染色的PI进行双染才能有效判定细胞凋亡。 PI（碘化丙啶，Propidium Iodide）是⼀种可对DNA染⾊的红色荧光染料，是⼀种溴化⼄啶的类似物，在嵌⼊双链DNA后释放红⾊荧光。尽管PI不能通过活细胞膜，但却能穿过死细胞破损的细胞膜⽽对核染⾊。因此，Oxazole yellow与PI联合使用，可直接用于细胞凋亡的检测，凋亡细胞呈现绿色荧光，死细胞同时呈现红色和绿色荧光阳性，活细胞很少或几乎没有荧光。
组分：
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使用方法（以流式细胞术为例）：
1. 细胞准备
（1）对于贴壁细胞，胰酶消化后用培养液重悬，并用预冷的 PBS 洗涤 1 次；胰酶消化时间不宜过长，以防引起假阳性。 注：用胰蛋白酶消化然后使细胞在最佳细胞培养条件和培养基中恢复约 30 min，然后染色。
（2）对于悬浮细胞，1000 rpm 离心 5 min，弃上清，用预冷的 PBS 洗涤 1 次。
2. 细胞染色
用预冷的 PBS 悬浮细胞，每个样品推荐的细胞用量为 106 个/mL，向 1 mL 的样品中加入 1 µL Oxazole yellow 和 1 µL
PI，轻轻吹打混匀。冰上避光孵育 30 min。 注：我们建议增加以下两组实验对照：
空白管：阴性对照组细胞，不加染料，用于调节电压。
单染管：阳性对照组细胞，只加 Oxazole yellow 和只加 PI 两管，用于调节补偿。
3. 流式检测
孵育完成后，可以直接进行流式细胞仪检测，也可以 1000 rpm 离心 5 min，吸弃上清，用 1 mL 预冷的 PBS 重悬样品后 进行流式细胞仪检测。Oxazole yellow 可以由 488 nm 激光激发，检测荧光发射光谱约在 530±30 nm 处（FITC 通道），PI 通 道发射光谱约在 617 nm处（PI 或 PE 通道）。
产品参数：
Oxazole yellow dye：Ex/Em = 491/509 nm（结合DNA）； Propidium Iodine：Ex/Em = 535/617 nm （结合D
注意事项：
1. 使用前请将产品瞬时离心至管底，再进行后续实验。
2. 荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。
3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
应用范围：
膜透性凋亡检测
Apoptosis refers to the cell autonomous and orderly death controlled by genes to maintain the stability of the internal environment. Apoptosis is different from cell necrosis. Apoptosis generally refers to a programmed cell death process that occurs during the development of body cells or under the action of some factors through the regulation of intracellular genes and their products. Cell necrosis is a cell death process that is caused by strong physical and chemical or biological factors to cause disordered changes in cells. The difference between apoptosis and necrosis lies in the characteristic morphological and biochemical changes, including the changes of cell membrane permeability and nuclear chromatin, the contraction of cytoplasm and the loss of membrane asymmetry. The oxazole yellow/pi membrane permeability apoptosis detection kit produced by our company is a dual fluorescence detection kit based on oxazole yellow and PI dyes. This kit is suitable for fluorescence microscopy, flow cytometry, fluorescence microplate reader and other fluorescence detection systems. Oxazole yellow is a non cell membrane penetrating cyanine monomer green fluorescent dye with high affinity for DNA. It basically has no fluorescence when it is not bound to DNA, but can emit bright green fluorescence after binding to DNA. When apoptosis occurs, the permeability of cell membrane changes. At this time, oxazole yellow can enter the cell and bind to DNA, emitting bright green fluorescence. Therefore, it is often used for the detection of apoptosis. It should be noted that oxazole yellow can also stain dead cells, so it needs to be double stained with PI that specifically fluorescently stains dead cells to effectively determine apoptosis. PI (propidium iodide) is a red fluorescent dye that can stain DNA. It is an analog of pyridine bromide that releases red fluorescence after embedding double stranded DNA. Although PI cannot pass through the membrane of living cells, it can cross the damaged cell membrane of dead cells to stain nuclei. Therefore, oxazole yellow combined with PI can be directly used for the detection of apoptosis. Apoptotic cells show green fluorescence, dead cells show both red and green fluorescence positive, and living cells have little or no fluorescence.
Components:
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Usage (using flow cytometry as an example):
1. Cell preparation
(1) For adherent cells, after trypsin digestion, resuspend in culture medium and wash once with pre cooled PBS; The digestion time of trypsin should not be too long to prevent false positives. Note: Digest with trypsin and allow the cells to recover in the optimal cell culture conditions and medium for about 30 minutes, then stain.
(2) For suspended cells, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and wash once with pre cooled PBS.
2. Cell staining
Suspend cells in pre cooled PBS, with a recommended cell count of 106 cells/mL per sample. Add 1 µ L Oxazole Yellow and 1 µ L to 1 mL of the sample
PI， Gently blow and mix well. Incubate on ice in the dark for 30 minutes. Note: We suggest adding the following two experimental controls:
Blank tube: negative control group cells, without dye, used to regulate voltage.
Single staining tube: Positive control group cells were treated with only two tubes, Oxazole yellow and PI, for regulating compensation.
3. Flow detection
After incubation, the sample can be directly detected by flow cytometry, or centrifuged at 1000 rpm for 5 minutes, the supernatant can be aspirated, and the sample can be resuspended in 1 mL of pre cooled PBS for flow cytometry detection. Oxazole yellow can be excited by a 488 nm laser, and the detected fluorescence emission spectrum is around 530 ± 30 nm (FITC channel), while the PI channel emission spectrum is around 617 nm (PI or PE channel).
Product parameters：
Oxazole yellow dye:ex/em = 491 / 509 nm (bound DNA); Propidium iodine:ex/em = 535 / 617 nm (combined with D
Matters needing attention：
1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments.
2. fluorescent dyes have quenching problems. Please try to avoid light to slow down fluorescence quenching.
3. for your safety and health, please wear experimental clothes and disposable gloves.
Scope of application：
Membrane permeability apoptosis assay
规格
英文名称
Oxazole yellow/pi membrane permeability apoptosis detection kit
储存与运输
储存条件
避光,-20°C储存
运输条件
超低温运输
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高级数据
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 B2517312 |
分析证书 | O598364 |
溶液计算器
常见问题
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