Q：使用范圍？

A：該試劑主要是檢測動物活細(xì)胞內(nèi)的活性氧水平，對于植物可以在制備原生質(zhì)體后進(jìn)行檢測使用，該試劑盒不能檢測體內(nèi)的活性氧試劑。

Q：如何稀釋？

A：無血清培養(yǎng)基、HBSS或是PBS等緩沖液進(jìn)行稀釋，稀釋后不穩(wěn)定，建議現(xiàn)配現(xiàn)用。

Q：探針顏色？

A：綠色熒光。

Q：.如何避免過高的熒光背景？

A：探針孵育后，一定要洗凈殘余的未進(jìn)入細(xì)胞內(nèi)的探針，再檢測熒光信號，否則會導(dǎo)致背景較高。

Q：探針一般孵育多長時間？

A：37℃細(xì)胞培養(yǎng)箱內(nèi)避光孵育30min。

Q：可以檢測組織嗎？

A：該試劑主要是檢測動物活細(xì)胞內(nèi)的活性氧水平，因而不能檢測組織切片，對于活體組織內(nèi)的檢測沒有驗證過，盡量參考文獻(xiàn)，如果是將活體組織分散成單細(xì)胞進(jìn)行檢測并保證細(xì)胞的活性也是可以的。

Q：檢測儀器？

A：熒光顯微鏡、共聚焦顯微鏡、酶標(biāo)儀、流式細(xì)胞儀。

Q：可以檢測正常細(xì)胞中的活性氧的含量嗎？

A：正常細(xì)胞中活性氧含量很低，該試劑無法檢測正常細(xì)胞中活性氧的含量。

Q：我用的是同一支探針，未分裝，前 5 次都可以染上顏色，而且熒光信號很好， 這次沒有染上顏色？

A：大致有以下3個原因：1.細(xì)胞狀態(tài)不好，導(dǎo)致染色效率低；2.陽性藥物誘導(dǎo)時間過短，一般 37℃ 避光孵育 30 min-4 h 可顯著看到活性氧水平提高；3.這個探針反復(fù)凍融 4 次以上，探針性能會不穩(wěn)定，染色效率降低，以及熒光信號不穩(wěn)定（時強時弱、易猝滅）。建議拿到探針后，分裝避光保存在-20 C 冰箱中，避免反復(fù)凍融。

Q：活性氧檢測試劑盒,這個試劑盒，我檢測出了熒光值，怎么作圖？熒光值和ROS值之間是否有換算公式？

A：是不需要換算成ROS值。橫坐標(biāo)應(yīng)該就是您實驗組和正常組。

Q：平均熒光強度怎么反應(yīng)ROS水平？

A：實驗組平均熒光強度與正常組相比高，而且有顯著性差異，就說明活性氧升高?；蛘吣昧魇綑z測的話是用峰圖表示。

Q：如果是酶標(biāo)儀檢測，測得的峰值一般怎么進(jìn)行對比？實驗組和對照組的熒光值？是作圖嗎？

A：實驗組平均熒光強度與正常組相比高，而且有顯著性差異，就說明活性氧升高。是做統(tǒng)計圖，一般為柱狀圖。

Q：那流式可以又熒光峰值的偏移對嗎？

A：是的，流式是用峰值的偏移來表示結(jié)果。

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