Hi,

Could you elaborate on the goal of your LC-MS experiment?

Do you, for example, want to identify as many proteins in your sample as possible? Or something else?

Some additional information would be helpful!

hi!!!!!!

I have secretory proteins in hbss after treatment to the protozoa. And i want to identify those secretory proteins withe help of LC-MS. mixture of several proteins are present in the hbss and i did tca - acetone for the purification and now i want to identify them with help of LC-MS,

Hi,

I think you are interested in secretome analysis of some protista with LC-MS/MS. I suggest you:

1. extract and concentrate proteins from culture medium with wide pore (about 300 A) C4 or DVB solid phase extraction cartriges. Dry in speedvac.

2. Perform in solution digestion using DOC method.

3. Analyse. If protozoan proteome is underrepresented in protein database, use transcriptome to predict ORFs, and use them as database for search. Use TransDecoder to predict ORFs from assembled transcriptome, and filter to 99% nonredundandcy with CD-HIT).

4. Good luck!