overexpression of a protein from a HA tagged plasmid - western blot detection (Feb/16/2007 )
I am trying to express a protein in human cells but cannot see by Western blot if I have expressed the protein. I have seen in some papers that people western blot for HA tag to show expression of the protein.....why? It doesn't make sense to me that HA could be detected but the protein cannot. Can someone explain this to me? I am quite cluless when it comes to anything molecular. Thanks in advance
Maybe it has to do with how your protein folds. Maybe the HA tag is accessible to the antibody whereas your protein epitope is not.
Sometimes one doesnot have good antibodies toward the protein of interest. therefore people construct genes with the HA tag. usually the tag doesnt interfere and upon expression in cells, the protien can be detected very efficiently due to the HA tag.
Its not only the HA tag , its for epitope tags in general. Like myc or FLAG tag, etc...