crisalis82

I do use a protease inhibitor. I get bands and I get them at the correct general MW but I can't get anything consistent.  Even when I get something that looks good there seems to be a larger margin of error that I would think when I quantify the relative concentration of my protein samples.  Is there anything else that might explain this?  I feel like my Westerns are otherwise great.

I was given a general protocol which I now think is insufficient and having been blaming myself for the weird results for near a year.  Does it make sense that the presence of a high concentration of lipids would result in inconsistent and inaccurate Western results?  If so, can anyone give me references that I can use when I try to explain this to my PI?  I should probably also explain that he is not a biologist so I don't really know the full depth of his background in regards to this particular procedure.

If my proteins are associated with lipid droplets, would the normal denaturing SDS PAGE prep separate them or could it be that some of my sample is just floating up out of the wells?