Common precautions, operation points and optimization methods
1.How to get as much exosome protein as possible for Western Blotting experiments?
a.Maximize the amount of initial sample used for exosome extraction;
b.Use ultracentrifugation, or a reliable exosome extraction kit (such as EZB-exo1, EZB-exo2, etc.) to ensure the extraction efficiency of exosomes;
2.If the content of exosomes is very little, how to adjust the amount of exosome protein extraction buffer to optimize the experiment?
For samples with little exosome content, as little as 60 pl of exosome protein extraction buffer can be added, shake well and mix thoroughly, and doing the subsequent steps.
3.What should I do if the experiment needs to quantify the protein concentration?
If the extracted sample needs to be used for protein quantification, it needs to be lysed with RIPA buffer (requires protease inhibitor), then mixed with 6x protein loading buffer, and then heated and denatured (For sample extracted from this kit, it is recommended to control the initial sample volume to ensure the same amount of sample. It is not recommended to measure the protein concentration).