Regarding the expression of phosphorylated c-Jun (p-c-Jun) in kidney tissue observed through Western blotting, even under high sensitivity settings and after 30 minutes, the detection quantity was low. The primary antibody was set at a dilution of 1:1000, and the secondary antibody was set at a dilution of 1:10,000. This is the first attempt at detecting p-c-Jun. How well can it be detected under these conditions?
Whatever you do, make sure you have a positive control and include phosphatase inhibitors in your lysis buffer. Jun phosphorylation is transient and occurs only under specific conditions. We use pjun abs from Cell signaling.
Sébastien Soubeyrand I also use pjun abs from Cell Signaling. This time, I didn't include a phosphatase inhibitor, so that might be the reason. Following your advice, I'm going to try Westernblot again. Thank you for your answer.
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