hi
Im using DHE to detect ROS in leukemia primary cells (mouse)
I noticed that even if the concentration of DHE is equal between well plates, if the cell number between different wells is not equal or at least comparable, the results are not consistent.
usually- the more the cells, the less ROS detected on FACS (MFI)
anyone knows about these relationships?
thank you
Avigail
Hi Avigail,
You need to analyze it under the same number of cells, such as 10,000 cells when using FACS. However, as you mentioned, the cell-cell interaction may be one of the reasons to affect the final ROS production. You need to seed the same number of cells among different wells.
Hi, in addition to cell number determination you can measure the concentration of total proteins. Normalizing your DHE values to total mg of proteins can provide increased accuracy of your measurements.
Regards, Christian
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