Hi
I am trying to develop positive control for protein, lipid and DNA as a measure of oxidative stress in formalin fixed paraffin embedded normal tissues during immunohistochemistry.
For protein damage, nitrotyrosylation of protein was produced by peroxynitrite and DNA damage induced by DNAse in formalin fixed paraffin embedded normal tissues.
However, I failed to produce lipid damage in formalin fixed paraffin embedded tissues by using Ferrous Sulphate and Hydrogen peroxide and Ferrous Sulphate and Hydrogen peroxide. I used antiMDA antibody (malondialdehyde antibody ab6463 from abcam) to detect lipid damage.
Does anyone have experience in producing lipid damage in formalin fixed paraffin embedded normal tissues?
Best regards
Debashish
I think your task and the methods belong to different centuries.
Did you heard about the Isoprostane Pathway of Lipid Peroxidation?
You get isoprostanes simply by the storage of your samples in the freezer at -20C. If you store a fish or fat chicken in the freezer you also damage lipids. O2 dissolves better at the cold temperature.
Good luck.
Alexander Panov
PS. To help you, I have attached the Manuscript of my paper, which I expect will be accepted soon. So far, it is only for your personal use (or other readers of this message}.
Thanks, Panov. Your article makes for an interesting reading. However, what I am trying to validate is a primary antibody against MDA, which was not detected in my normal samples (formalin fixed and paraffin embedded). In the absence of any positive control tissues, I am forced to prepare my own positive control by the method chosen.
However, I would like to try the method suggested by Suji. Thanks, Suji.
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