Basically, I am trying to extract total proteins (all the possible type of proteins including soluble/cytosolic and membrane-associated proteins or may be the protein is expressed inside the chloroplast), can anyone please suggest what type of extraction buffer should be used for the same.
Thanks.
As Ali wrote, you will never isolate all proteins with one buffer. Some proteins will require high pH, some low pH, some lots of salt, some none of salt.
At least you can include detergent. That will help to break all the membranes and release all proteins. But again, some proteins will favour neutral detergents, some zwitterionic etc.
Thank you Dr. Ali and Dr. Tomas. We are not looking for active proteins, we just want to see if the protein is expressed. We are interested in cytosolic/ soluble as well as chloroplast envelope proteins, can you please suggest us an effective buffer/detergent for the same.
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