3 replies to this topic

[eman]

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

[protolder]

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

[BioMiha]

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...

[Allenwood]

Nice information........

thanks.