产品说明书
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已发表文献
产品简介
MitoSOX Red Mitochondrial Superoxide Indicator，一种新型线粒体荧光探针，可特异性靶向线粒体，从而选择性检测线粒体内的超氧化物。该探针可透过活细胞膜，并选择性进入线粒体。一旦进入线粒体，该探针即可被超氧化物氧化发出红色荧光。本品可被超氧化物而非其他活性氧类(ROS)和活性氮类(RNS)快速氧化。氧化产物结合核酸后，可产生大量荧光。
本品以粉末形式提供。
产品性质
|
分子式（Formula） |
C43H43N3IP |
|
分子量（Molecular Weight） |
759.71 |
|
荧光光谱 （Fluorescence Spectral） |
探针本身的Ex/Em= 356/411 nm；氧化型MitoSOX探针-核酸的Ex/Em= 396/580 nm |
|
溶解性（Solubility） |
溶于DMSO |
|
纯度（Purity） |
≥97% |
|
结构式（Structure） |
|
储存条件
-25 ~ -15°C避光干燥保存，避免反复冻融，有效期2年。
使用说明
1. 储存液制备（5 mM）
本品是以粉末形式提供，使用前需将本品回温至室温。
向50 µg MitoSOX Red Mitochondrial Superoxide Indicator中加入13 µL DMSO，混匀即可配制成5 mM的储存液。储存液可按照单次用量分装，在-20℃避光保存，避免反复冻融。
2. 操作方法
不同的实验目的使用不同的探针浓度，以下的起始操作条件仅作参考，可根据细胞类型和其他的相关因素进行适当调整。
1）配置500 nM工作液。（将5 μl的储存液添加至50 mL含Ca2+、Mg2+的HBSS缓冲液）
注：对于不同的细胞类型，将染色溶液浓度优化在100 nM至1 µM之间，以最大限度地提高信噪比并最大限度地降低细胞毒性。
2）加载细胞：加入1 - 2 mL探针工作液充分覆盖爬片生长的细胞，于37°C、5%CO2下避光孵育30 min。
3）清洗：用预热的合适缓冲液轻洗细胞3次。
4）镜检：选择合适的复染液对细胞进行复染，封片后进行观察。
注意事项
1. 本产品仅作科研用途。
2. 为了您的安全和健康，请穿实验服并佩戴一次性手套操作。
3. 本品易被氧化，避免将本品与空气接触。
4. 本品属于溴化乙锭（EB）衍生物，具有毒性，请小心操作。
Ver.CN20241124
Q：是可以用来做免疫荧光的吧？是可以用荧光显微镜看的对吧？
A：可以用荧光显微镜，客户的实验目的如果是检测活细胞线粒体超氧化物，就可以用这个的，这个和免疫荧光是没有关系的，免疫荧光是杂抗体的。
Q：之前买的这个探针效果不好，入核了。有没有方法或者范图呢？有推荐浓度么？
A：这个没有效果图，可降低浓度试一下，这个样本不一样浓度也不一样，需要客户参考一下文献。
Q：我想用线粒体 mito-green 和 mitosox 混染细胞，如果染的话，两个染料能不能同时混合着染，还是按照一个先后顺序来染？
A：建议分先后顺序，先染一个洗完后，再染另一个。
Q：探针溶解方法可以用其他代替吗
A：可用PBS 溶解。
Q：40778ES50 可以做流式检测吗？
A：可以。
Q：染色效果不好，如果提高染色效率？
A：缓冲液使用HBSS（含钙镁）培工作液进行染色。
Q：MitoSox Red 线粒体超氧化物红色荧光探针激发波长是否可以选择396 nm？
A：可以的。虽然该产品在510 nm激发时会显示较强荧光信号，但据发现在396 nm超氧化物产物能被选择性激发且由其它非特异性氧化产物造成的干扰最小，所以激发波长选择396nm完全可以，这会使得检测结果更准确。参考文献PMID: 17015830。
Q：染色工作液体积应该加入多少？染色时间可以延长吗？
A：对于35mm共聚焦小皿，推荐1-2ml染色液；对于96孔板推荐100ul染色液。推荐37℃孵育10min，但可适当延长孵育时间到10-20min，最长建议不超过30min。
Q：染色时出现入核情况该怎么处理？工作浓度是否可以调整？
A：染色时如遇明显入核情况，可以通过降低工作浓度以及缩短染色时间来进行调整。对于工作浓度，如出现染色入核及过曝可降低为100 nM到1 μM；如遇染色信号较弱也可调整高于5 μM，同时需对染色工作液进行预热。
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