cytoplasmic protein - (Nov/25/2013 )
I recemy;y extracted cytoplasmic protein from a 10cm dish using the NE-PER (R) Nuclear and Cytoplasmic Extraction kit (78833 ).
However, my protein is too concentrated to run for western. I need to run 5ug-10ug for western which is at the moment equivalent to 1ul of my concentrated sample. This is too concentrated for running western. How can I dilute the protein? I was thinking of adding some additional cytoplasmic extraction buffer that came with the Kit?
Having protein that is too concentrated is a great problem to have. You can load 1uL on a gel if you have the right kind of syringe. Otherwise, I would dilute the sample in 1X sample buffer just prior to loading the gel - no need to add extra salts, buffers, etc. that are likely in the extraction buffer to the sample.