His-tag purification and his-antibody for western
Posted 10 January 2002 - 01:48 AM
I met a problem in purification my protein expressed from pichia KM71 cell. it is a novel membrane protein and I can detect the expression in the crude membrane of km71. After purification with the Qiagen Ni-NTA kit, I got a very sharp band, a little smaller than the expected size. To my surprise, the band can not be detected with anti-His in Western. WHY??
Hoping get some information from any one.
Posted 13 February 2002 - 09:52 AM
I think, may be, that HIS sequence could be eliminate during expression.
You can see protein after purification with coomassie but not detected with anti-HIS antibody.
Posted 12 November 2003 - 07:37 PM
Edited by volkandemir, 12 November 2003 - 07:37 PM.