Protein extraction - Western Blot
I have a doubt regarding protein extraction for Western Blot. I'm using the same protein samples that I used for ELISA and for ELISA I used 1 ml of PBS and 10 µl of halt protease inhibitor to grind the brain followed by high speed centrifugation at 4˚C. I used this method for ELISA and it worked.
As I had a lot of protein extract left, I used it for Western Blots. I tried doing it with TLR9 antibody and I saw the specific bands. I also ran a Coomassie staining to see if there is a protein degradation but found that proteins were good.
My question is: Will there be any problem when I publish as I haven't used the RIPA buffer to lyse the hippocampi and instead used a PBS + Halt protease inhibitor combination?