Protocol Online logo
Top : New Forum Archives (2009-): : Protein Expression and Purification

protein purification - (Dec/21/2010 )

i want to ask about how can i make purification to my protein? the M.wt of the protein is 5kDa in expression vector PET-29 in Bl21 bacteria


Hola, in pET 29 you can have your protein fused with Nt6His,-is your case?.If yes, after lysis of bacterial pellet I would use any of the commercial resins of Ni++ or Co++ with th addecuate device for the volume of culture, spin columns, FPLC colums, or resin in batch under the conditions of the supplier. Buena suerte


If you are expressing your protein in E. coli the best way to purify it is by attaching a His tag (or any other tag). Otherwise you can use a number of methods depending on the characteristics of your protein. There is no pre-defined way to purify a protein, it all depends on what the protein is like, what the sample is like, how clean you need it to be, what you need it for, etc., etc...


Nice information........