A common negative control setup in immunofluorescence is the omission of primary antibodies.
What purpose does a negative control serve in these contexts? Is it just to get an idea of the background from secondary antibodies - or is it important in experimental design?
Hi,
it shows you the specificity of your secondary Antibody and with this shallmake clear that your signal is coming solely from the detected target of the primary Ab. You can also use primary control IgGs
Hello Austin Morrissey
The goal of a negative control is to demonstrate that the reaction visualized is due to the interaction of the epitope of the target molecule and the paratope of the antibody.
The negative control setup in immunofluorescence which is the omission of primary antibody is a control for nonspecific binding of the secondary antibody.
This result is not evidence for the specificity of staining with the primary antibody.
The proper negative control would be substitution of serum or isotype-specific immunoglobulins at the same protein concentration as the primary antibody.
Best.