MitoSOX Red Mitochondrial Superoxide Indicator 線粒體超氧化物紅色熒光探針
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產(chǎn)品簡介
MitoSOX Red Mitochondrial Superoxide Indicator，一種新型線粒體熒光探針，可特異性靶向線粒體，從而選擇性檢測線粒體內(nèi)的超氧化物。該探針可透過活細(xì)胞膜，并選擇性進(jìn)入線粒體。一旦進(jìn)入線粒體，該探針即可被超氧化物氧化發(fā)出紅色熒光。本品可被超氧化物而非其他活性氧類(ROS)和活性氮類(RNS)快速氧化。氧化產(chǎn)物結(jié)合核酸后，可產(chǎn)生大量熒光。
本品以粉末形式提供。
組分信息
|
組分名稱 |
40778ES50 |
|
MitoSOX Red Mitochondrial Superoxide Indicator 線粒體超氧化物紅色熒光探針 |
50 μg |
產(chǎn)品性質(zhì)
|
分子式（Formula） |
C43H43N3IP |
|
分子量（Molecular Weight） |
759.71 |
|
熒光光譜 （Fluorescence Spectral） |
Ex/Em= 396/610 nm |
|
溶解性（Solubility） |
溶于DMSO |
|
純度（Purity） |
≥97% |
|
結(jié)構(gòu)式（Structure） |
|
儲(chǔ)存條件
-25 ~ -15°C避光干燥保存，避免反復(fù)凍融，有效期2年。
使用說明
1. 儲(chǔ)存液制備（5 mM）
本品是以粉末形式提供，使用前需將本品回溫至室溫。
向50 µg MitoSOX Red Mitochondrial Superoxide Indicator中加入13 µL DMSO，混勻即可配制成5 mM的儲(chǔ)存液。儲(chǔ)存液可按照單次用量分裝，在-20℃避光保存，避免反復(fù)凍融。
2. 操作方法
不同的實(shí)驗(yàn)?zāi)康氖褂貌煌奶结槤舛?，以下的起始操作條件僅作參考，可根據(jù)細(xì)胞類型和其他的相關(guān)因素進(jìn)行適當(dāng)調(diào)整。
1）配置500 nM工作液。（將5 μl的儲(chǔ)存液添加至50 mL含Ca2+、Mg2+的HBSS緩沖液）
注：對(duì)于不同的細(xì)胞類型，將染色溶液濃度優(yōu)化在100 nM至1 µM之間，以最大限度地提高信噪比并最大限度地降低細(xì)胞毒性。
2）加載細(xì)胞：加入1 - 2 mL探針工作液充分覆蓋爬片生長的細(xì)胞，于37°C、5%CO2下避光孵育30 min。
3）清洗：用預(yù)熱的合適緩沖液輕洗細(xì)胞3次。
4）鏡檢：用熒光顯微鏡或細(xì)胞流式儀進(jìn)行檢測分析。
注意事項(xiàng)
1. 本產(chǎn)品僅作科研用途。
2. 為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并佩戴一次性手套操作。
3. 本品易被氧化，避免將本品與空氣接觸。
4. 本品屬于溴化乙錠（EB）衍生物，具有毒性，請(qǐng)小心操作。
Ver.CN20241224
Q：是可以用來做免疫熒光的吧？是可以用熒光顯微鏡看的對(duì)吧？
A：可以用熒光顯微鏡，客戶的實(shí)驗(yàn)?zāi)康娜绻菣z測活細(xì)胞線粒體超氧化物，就可以用這個(gè)的，這個(gè)和免疫熒光是沒有關(guān)系的，免疫熒光是雜抗體的。
Q：之前買的這個(gè)探針效果不好，入核了。有沒有方法或者范圖呢？有推薦濃度么？
A：這個(gè)沒有效果圖，可降低濃度試一下，這個(gè)樣本不一樣濃度也不一樣，需要客戶參考一下文獻(xiàn)。
Q：我想用線粒體 mito-green 和 mitosox 混染細(xì)胞，如果染的話，兩個(gè)染料能不能同時(shí)混合著染，還是按照一個(gè)先后順序來染？
A：建議分先后順序，先染一個(gè)洗完后，再染另一個(gè)。
Q：探針溶解方法可以用其他代替嗎
A：可用PBS 溶解。
Q：40778ES50 可以做流式檢測嗎？
A：可以。
Q：染色效果不好，如果提高染色效率？
A：緩沖液使用HBSS（含鈣鎂）培工作液進(jìn)行染色。
Q：MitoSox Red 線粒體超氧化物紅色熒光探針激發(fā)波長是否可以選擇396 nm？
A：可以的。雖然該產(chǎn)品在510 nm激發(fā)時(shí)會(huì)顯示較強(qiáng)熒光信號(hào)，但據(jù)發(fā)現(xiàn)在396 nm超氧化物產(chǎn)物能被選擇性激發(fā)且由其它非特異性氧化產(chǎn)物造成的干擾最小，所以激發(fā)波長選擇396nm完全可以，這會(huì)使得檢測結(jié)果更準(zhǔn)確。參考文獻(xiàn)PMID: 17015830。
Q：染色工作液體積應(yīng)該加入多少？染色時(shí)間可以延長嗎？
A：對(duì)于35mm共聚焦小皿，推薦1-2ml染色液；對(duì)于96孔板推薦100ul染色液。推薦37℃孵育10min，但可適當(dāng)延長孵育時(shí)間到10-20min，最長建議不超過30min。
Q：染色時(shí)出現(xiàn)入核情況該怎么處理？工作濃度是否可以調(diào)整？
A：染色時(shí)如遇明顯入核情況，可以通過降低工作濃度以及縮短染色時(shí)間來進(jìn)行調(diào)整。對(duì)于工作濃度，如出現(xiàn)染色入核及過曝可降低為100 nM到1 μM；如遇染色信號(hào)較弱也可調(diào)整高于5 μM，同時(shí)需對(duì)染色工作液進(jìn)行預(yù)熱。
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