4 replies to this topic

[FRAROAR]

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

[Vini]

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...

[Curtis]

how can you add 6 His?

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

[Vini]

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!  

[T C]

Hey,

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

Best,
TC