Several things might have gone wrong. You might have mistakenly placed the gel/membrane sandwich in the wrong orientation in the transfer cell, or the Pounceau stain might not have worked well. I'd start troubleshooting by first running a new gel and staining it with Coomassie to see if there is any protein actually in your samples.

agree with @ Alejandro Martin

regards

Check this link is useful

https://www.sigmaaldrich.com/technical-documents/protocols/biology/extraction-from-tissue.html

regards

Thank you for your answers. I will try with Coomassie as suggested by Alejandro to be sure if there was any protein in the gel or no.

Hi,

I increase the quantity of protein that I loaded (50 microgram instead of 30) and I did the Coomassie stain. I get a few band (4 bands) with a few intensity compared to a positive control (homogenate extract with RIPA buffer). Any suggestions!!!!

Thank you very much

you can run two same gels which one for starting and another one for west. Make sure you have protein.

In this case the Trizol procedure is not working as well as you expected. If you can switch to RIPA do so, otherwise I'd suggest you precipitate your samples with whatever method you prefer and resuspend them in a smaller volume, so that you can load more on SDS-PAGE.

Trizol is a known reagent for the extraction of nucleic acids and not proteins. May I suggest you use a buffer, contacting both ionic and non-ionic detergents, to extract proteins from your sample. Such an approach to extracting total proteins from both tissue and cells is well established.