I am trying to learn about this protocol in order to isolate proteins from plants material. Thanks in advance.
For little quantities you can use Immunoprecipitation, but I do not have experience in this field. This technique uses magnetic separation.
To isolate plant proteins you can use TCA/Acetone precipitation method. I am attaching one paper that could help you in this regard.
There are different methods for isolation of protein from plants one could be by isoelectric precipitation. Following steps could be followed for extraction by this method.
1. Remove the undesirable fractions eg. carbohydrates and fat or oil (if you are extracting from any oil seed). This will concentrate the protein in remaining portion.
2. Now dissolve that portion in the buffer of a particular pH (depending on the kind of protein you are targeting)
3. Then lower the pH to isoelectric point of protein, so that it can get precipitated.
4. Collect the protein precipitates by centrifugation.
5. These ppts. can be be further purified by dialysis.
Protein extraction method would depend on your application...I mean why you are isolating the proteins? If you just want to do SDS-PAGE or 2DE, use either TCA-Acetone or phenol method and in case you want to do some kind of enzymatic activities, use a Tris, HEPES or phosphate based extraction buffer.
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