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cytoplasmic protein


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#1 SF_HK

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Posted 25 November 2013 - 01:08 AM

I recemy;y extracted cytoplasmic protein from a 10cm dish using the NE-PER ® 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



#2 dimensionsbio

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Posted 25 November 2013 - 10:36 AM

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.






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