Posted 21 October 2009 - 10:49 AM
I have HA tagged my protein. My sequencing reaction looks good. Everything should be in frame.
I have added a single HA tag.
The problem is that my protein is expressed (by Western) but I cannot detect it with an HA tag antibody (I use a 3x HA tag for control).
Posted 13 November 2009 - 12:58 PM
Posted 15 November 2009 - 04:01 PM
My single HA tag works just fine, I am using a Sigma anti-HA rabbit antibody, which shows a strong band of the right size and a much weaker and slightly smaller band in the controls and treatments.
Is there a spacer between the HA and the protein of interest? If not, direct linkage has been known to affect the binding of antibodies to tags.
Edited by bob1, 15 November 2009 - 04:01 PM.