Hello,
I was wondering if anyone had ever performed a Dot Blot of Chlamydomonas
whole cell extract directly on PVDF or nitrocellulose membrane. I have a
175 kDa protein and I am not seeing anything when I do a western transferred
from a polyacrylimide gel. My concern about the dot blot on whole cell
extract is the pseudo-peroxidase activity of unbound heme. (My protein has
a very low expression and I am using an HRP conjugated secondary antibody.)
Does anyone know if there is perhaps something like a chelator that I could
add to the protein to precipitate out the heme? (I am going to try a pull
down, but if possible, I would like to see my protein without this step for
future experiments.)
Thank you in advance!
- Sara
