immunization w/ 6xHis-tagged protein

Tyson Graber tysong at home.com
Tue Sep 28 22:56:35 EST 1999


If you're thinking of immunizing w/ 150mM Imidazole...it probably won't be
toxic (short-term) to the rabbit using small amounts.....you should remove
as much of it as possible.  If you have several rabbits available you can
try a couple of things:

1.  EtOH precipitate the protein & re-suspend in PBS

2.  EtOH ppt, R/S in hydrolysed NaOH and neutralise with Acetic Acid,
followed by emulsification in Freund's Adjuvant (complete)

good luck


Takahiro Ito <ff87105 at hongo.ecc.u-tokyo.ac.jp> wrote in message
news:199909250648.PAA25722 at hongo.ecc.u-tokyo.ac.jp...
| Dear all,
| I am going to immunize a rabbit with hexa-His-tagged recombinant protein
| eluted with imidazole (150mM).
| Some said that imidazole is toxic to animals, so it should be removed from
| the fractions before injection. Could anyone tell me whether I should
remove
| imidazole or not? Any suggestions would be appreciated.
| Thank you in advance.
|
|





More information about the Methods mailing list