One Antibody in Western Blot - (Jun/26/2006 )
Is it possible to use only one antibody in western blot instead of two (primary and secondary)?
if you have a primary conjugated to a detector such as HRp then it should be fine, but the two antibody system allows greater specificity and sensitivity.
Thanks Bob! Does anyone use this method (one antibody)?
I've used a primary HRP-conjugated and it worked, but it depends on the Ab
You can also use fluorophore-conjugated primary antibodies. These are the sorts of molecules you would use in immunohistochemistry to visualise the distribution of specific proteins in tissue samples.
However, if it is for Western blots then everyone is correct. You will have massively reduced sensitivity due to the lack of amplification.