Looking for precipitation method to reduce the volume of my protein eluate

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AllenChiu
AllenChiu's picture
Looking for precipitation method to reduce the volume of my protein eluate

I want to reduce the volume of my eluate of protein complex by precipitation and resloving in small volume so I can load them into SDS-PAGE gel.The eluate comes from the elution of Strep-Tactin Superflow resin with elution buffer with 2.5mM D-Desthiobiotin in it.
The precipitation method should be able to precipitate each proteins in the former complex no matter the abundance is low or high.
The precipitation should be able to readily been resolved again without changing the profile of former composition of proteins.
Then what is your suggestion? As the TCA precipitation is hard to be resolved again.

PS:My overall purpose is to discover new interacting protein of protein of interest by Strep-Flag tandem purification.

Chin Fen Teo
Chin Fen Teo's picture
 Hi AllenChiu,

 Hi AllenChiu,

As for the method for protein precipitation, please look at an old post awhile back where you can find a paper that compared different method with 2D-gel.

You may pick any of the method that you think is easier with what you have in hands, but I personal favorite is to use the methanol chloroform method, as it is super fast compare to many other methods that require incubations... 

After performing the precipitation, if you boiled your sample directly in the Laemmli sample buffer, it should be able to get most of the stuffs back into solution.

Good luck.