I have been working with a tyrosine kinase inhibitor, which showed significant positive effect by declining ROS generated by TNF-a (10ng/ml, 1hr) in HUVEC.
I am using 20uM of DCFDA +plus compound (10nM, 100nM) for 30min and after washing (2times with PBS) treating TNF-a along with the same concentration of my compound. This gives considerably fine result.
But while doing a quenching study, I found that the compound has quenching action over the control (10uM H2O2).
I was wondering if I really need to consider the DCFDA concentration or there are other factors working here?
Thanks in advance.
There are those who would argue against using DCFDA at all. There is a new review published in Circ Res on measurement of ROS in the cardiovascular system which should be worth a look.
http://www.ncbi.nlm.nih.gov/pubmed/27418630
Dear David,
Thank you very much. But unfortunately it seems that we do not have access to that article. Could you please send me the article. I'll be much obliged.
Its really very very important for me to fix the problems I am having.
Best,
Taufiq
Having difficulties uploading for some reason, but they state clearly that they do not recommend use of DCFDA. Do you have access to EPR?
Dear David,
No problem. Got the article. Many other authors also do not recommend DCFDA, but unfortunately at present my lab is dealing with DCFDA only. And we don't have EPR facility.
That i was thinking lately. Thank you for uplifting. Since managing technical support has been a problem i would better be replacing dcfda. I'll let you know the whole thing Dear David.
appreciate your kind help, always