I have purified a recombinant protein and want to assay the protein. I have an idea to immunoprecipitate the protein and further pull down the protein native and subject it to PTM. I would like to know if this method works out or if there are other methods to detect the protein?
It would be good if you can mention how did you purify the recombinant protein which makes it easy to suggest how to detect the PTMs
It depends on what kind of ptm you want to analyze. If you have an idea what PTM you are expecting, you can directly do WB for detection; if you want to identify novel PTMs or have no ideas what PTMs this protein gonna have, you can identify them with mass spectrometry, usually requires enrichment of modified protein/peptides prior to MS.
If I am not wrong there is no ptm modifications in bacteria, since they do not have organelles required for ptm. most of the recombinant proteins expressed in bacteria do not have ptm's, to get a recombinant protein with correct ptm's mammlian cell or insect cell expression systems are used
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