Hi, I want to evaluate oxidative stress in a neuroblastoma cell line, I know about SOD1, but I think it's not enough, so I was investigating and I found 4-hydroxynonenal, I have this in my lab 4-HNE B5605, but I don't find the concentration to use. Also I don't think it is the best marker for inmunofluorescence, It is more used in western blot . Do you know other types of marker that I can use for oxidative stress in a cell culture? I will appreciate your help.
You could try using H2DCFDA. It is mainly used in fluorometry and flow cytometry but you can also find publications using it in immunofluorescence. I could provide you with a protocol but you would have to wait some weeks cause I am currently optimizing it. Use live cells, do a quick test using H2O2 as a positive control and I hope it works out for you.
I wish you best of luck,
George
Hello Ana Palacios Ochoa
You may use Malondialdehyde which is a product of lipid peroxidation and has been used as a biomarker of oxidative stress. Please see the link below for the Malondialdehyde antibody having IF application.
https://www.stressmarq.com/products/antibodies/monoclonal-antibodies/malondialdehyde-antibody-smc-515/?v=3e8d115eb4b3
You may also add Glutathione Peroxidase 1 to your list. In the critical redox system of mammalian cells, glutathione peroxidase (GPX) is the most prominent family of proteins with a multifaceted function that affects almost all cellular processes. They have long been used as antioxidant enzymes to play an important role in combating oxidative stress and maintaining redox balance. Please see the link below for the Glutathione Peroxidase 1 antibody having IF application.
https://www.abcam.com/products/primary-antibodies/glutathione-peroxidase-1-antibody-ab22604.html
Best.
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