Hi,
I am working on a membrane bound protein, but due to experimental approach of checking its activity I cannot solubilize my protein and thus I do not use any detergent in my lysis buffer. However I need to quantify my protein by western blot and I see my protein band on membrane but I encounter with problems like smiley band, smearing etc. and not clear band as I encounter with problem in loading the protein in gel too due to its thickness or non- homogeneous mixture.
Could anyone suggest what can I do to improve the difficulties I am facing.
Thanks.