How can i optimize the protein amount obtained in cardiac homogenates when working with limited amounts of tissue (mice)? (Homogenates will be used to measure oxidative stress biomarkers)
Maiara Providello you can calculate the total protein concentration by using BCA.
I think this research may be help you: https://dmsjournal.biomedcentral.com/articles/10.1186/1758-5996-3-17
Spectrophotometric methods to optimize the amount of protein have been investigated in the following article
https://www.sciencedirect.com/science/article/abs/pii/S0026265X00000175
To homogenize add 500ul cold PBS to pH 7.4, Centrifuge for 20 minutes at full speed, take the supernatant and carry out protein determination by BCA and another for oxidative stress
:)
For stress marker studies, you prepare 10 percent tissue homogenate. After centrifugation, supernatant will be used for protein estimation (Lowery's or BCA method) and stress marker studies.. Once you done with stress marker estimation, you need to optimize with protein content (eg. n moles or u moles of aaaa/mg of protein)... I hope, my research article may be help you..
https://www.sciencedirect.com/science/article/abs/pii/S0891584919308494
Exactly, for example if you determine NO, it is necessary to correlate the uM NO with mg of protein obtained
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