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His Tag protein - (May/18/2009 )

Good morning dear all, I want to espress a big protein..about 1070 aa.
I want to insert six His to this protein to improve the purification. are suffient 6 His or the protein is to big to purify with a Ni column?

Thanks,

Fra

-FRAROAR-

FRAROAR on May 18 2009, 04:54 AM said:

Good morning dear all, I want to espress a big protein..about 1070 aa.
I want to insert six His to this protein to improve the purification. are suffient 6 His or the protein is to big to purify with a Ni column?

Thanks,

Fra



that should not be a problem. 6 His residues are sufficient...

-DRN-

how can you add 6 His?

why not use a commercially available his-tag vector. what would be the difference?

-Curtis-

Curtis on May 18 2009, 07:01 AM said:

how can you add 6 His?

why not use a commercially available his-tag vector. what would be the difference?



i guess FRA meant using a his-tag vector only, just simplified the sentence! B)

-DRN-

Hey,

Prefer a Cter His tag as that will ensure that no truncated forms of the proein purify.

Best,
TC

-T C-