We are conducting experiments with antioxidant supplementation in zebrafish and we will like to evaluate some mitochondrial parameters (membrane potential, ROS generation, swelling assays, etc) in liver. A protocol and the liver mass necessary for mitochondria isolation should be welcomed.
Thank you very much in advance.
Jose
Dear Dr Monserrat
Sorry I have no experience about the mitochondria isolation! Please let me know if you need others information
With best regards
Jamel JEBALI
Dear José Maria,
You need to excise the liver from zebrafish, suspend it in isolation medium and homogenize it, and subject this homogenate to differential centrifugation (subcellular fractionation) in order to isolate the mitochondrial fraction (containing functional mitochondria. You may want to refer to the series, Methods in Enzymology. The following would help...
http://www.nature.com/nprot/journal/v2/n2/fig_tab/nprot.2006.478_F4.html
http://www.cshlpress.com/pdf/sample/2014/subcellfraction/SubFractCH13.pdf
Thank you everybody for their useful and valuable suggestions.
José
dear José Maria Monserrat , did you have success in isolating zebrafish mitochondria? If you did, can you share the protocol for it?
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