I am doing westerns with human brain tissue and the westerns seem to be working fine. I'm getting a nice band of the right weight, but nothing shows up when I ponceau stain.
But here's the thing: I am using revived fixed tissue. Frozen tissue from the same case stains with ponceau.
Any thoughts? Is it okay that it blots just fine but doesn't ponceau?
I coimpletely agree. It might be a function of you using revived fixed tissue that Ponceau does not yiled obvious staining but that being so is academic. If the ladder transfers as already mentioned sand you are seeing the correct band then in principle your Western blot is valid
Some prople dont even bother using ponceau; Just look at ladder transfer as a proxy for efficient sample lysate transfer
SDS might interfere with Ponceau staining. Washing thoroughly the membranes could help. But anyway, if you have the bands and the ladder, it means that the proteins have been succesfully transferred and the Ponceau is not working properly.
I am using nitrocellulose. Is PVDF better to use with Ponceau?
Thanks everyone for your answers! They were all really helpful. I was just questioning the validity of my westerns because of the lack of Ponceau. But now I'm not worried, thanks!
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