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protein purification in acanthamoeba lysate - (Jul/28/2013 )

hi I am still new to protein purification
so i would like to hear your purify unknown protein from acanthamoeba lysate  should i use fplc or hplc .....
which one the best.....
thank you.....biggrin.png ...


the best purification depends on the protein and what you know about it (is it tagged? an enzyme? structural protein? etc).


hplc and fplc are similar except hplc generally uses higher pressure and less sample. fplc may be more for preparative purposes (not necessarily).


how do you detect and "identify" the unknown protein?


thank you for reply my massage...... 


i will use sds page and send the data for analysis.


did you have any other method for unknown protein determination.


thank you..biggrin.png


you can cut the band out of the gel and analyze the sequence with lc-ms or transfer to pvdf and perform edman degradation sequencing. then compare to a protein database.