Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

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?

 

Many thanks

-SF_HK-

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.

-dimensionsbio-