What kind of problem may face without isolation ? Usually after homogenization the liver, the microsomal part is separated and used for EROD assay?
Dear Md Saiful Islam,
The isolation of microsomal fractions is a mandatory step. As you know, the induction of cytochrome P450 monooxygenase 1A (CYP1A) is most commonly measured in terms of EROD ( 7-ethoxyresorufin deethylation) activity since this is a very sensitive agent. However, and since ethoxyresorufin-O-deethylase (EROD) can react with other entities (proteins) the measurement using this assay will be incorrect if the microsomal proteins is not purified according to the protocol for EROD.
Hoping this will be helpful,
Rafik
Dear Professor Rafik Karaman
Thank you so much for your informative reply. I understand the possible sources of interference.
However, is there any opportunity to combine the electrochemical measurement with fluorometry for EROD assay?
Thank you.
Dear Md Saiful Islam,
It is a very good question. I think the answer is yes.
Please read the recent publication contained in the following link:
http://pubs.acs.org/doi/pdf/10.1021/ac5044362
Hoping this will be helpful,
Rafik
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