Amount of Cell Lysate for Western Blot - (Feb/19/2008 )
What is the lowest amount of cell lysate that can be reliably used on a typical Western blot? I have been loading 50-75 ug per lane in the past. However, I now need to do Westerns on lower amounts of sample. Will 10-20 ug work?
I use normally 20-30 µg, that is absolutely no problem for the most proteins. In case that your on is only at the lowest level expressed, you may need to use more. But 25 µg for starting is good.
I frequently load 3,5 ug. So 10-20 shouldn't be a problem!