Get a positive control Treat your cells with Oligomycin or Antimycin These mitochondrial inhibitors should dramatically increase ROS generation.
x*a
12 楼
最好不要算得那么准确,最好提前几天
B*M
13 楼
该怎样证明一个不存在的东西不存在呢?
【在 o****b 的大作中提到】 : : 但没有佛法, 你又如何证明轮回存不存在呢?
m*d
14 楼
Can I use the H2O2 as my positive control?
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【在 h*******o 的大作中提到】 : Get a positive control : Treat your cells with Oligomycin or Antimycin : These mitochondrial inhibitors should dramatically increase ROS generation.
I don't recommend H2O2, as the reagent itself also elicits DCF fluorescent signal. Then it will be too hard to distinguish between ROS generated by the cells and ROS attributed to H2O2. Also, you might want to consider MtSOX for your assay.
【在 m**********d 的大作中提到】 : Can I use the H2O2 as my positive control? : : .
That's a very good suggestion, thanks! But I still have a minor question about this DCF chemical: According to the manual, it is permeable through cellular membranes until it is cleaved of the ester group and becomes trapped in the cell, where it is hydrolyzed by cellular ROS. Considering that it only reacts with intracellular ROS, plus I wash thoroughly after DCF loading to get rid of extracellular hydrolyzed DCF, if there's any, then H2O2 seems to be a good positive control. Looking forward to hear about your opinions
the cells and ROS attributed to H2O2.
【在 h*******o 的大作中提到】 : I don't recommend H2O2, as the reagent itself also elicits DCF fluorescent : signal. Then it will be too hard to distinguish between ROS generated by the cells and ROS attributed to H2O2. : Also, you might want to consider MtSOX for your assay.
H2O2 is also cellular membrane permeable. Unless you treated cells with H2O2, washed it off and waited for sufficient amount of time, it is possible that exogenous H2O2 will interfere with your assay. I
it is
【在 m**********d 的大作中提到】 : That's a very good suggestion, thanks! But I still have a minor question : about this DCF chemical: : According to the manual, it is permeable through cellular membranes until it : is cleaved of the ester group and becomes trapped in the cell, where it is : hydrolyzed by cellular ROS. Considering that it only reacts with : intracellular ROS, plus I wash thoroughly after DCF loading to get rid of : extracellular hydrolyzed DCF, if there's any, then H2O2 seems to be a good : positive control. : Looking forward to hear about your opinions :