In a Flow Cytrometry, one can add an Isotype Control Antibody to allow another a

Question

In a Flow Cytrometry, one can add an Isotype Control Antibody to allow another antibody to bind more specific to the cells.

My question is, how can the Isotype Control Antibody add specificity to the other antibody? My thought was, that the Isotype Control Antibody binds to all the antibody receptors, but with a "weak" binding. When the other antibody (e.g. CD73) tries to bind, it can make a stronger binding with the CD73 antibody receptor, kicking the Isotype Control Antibody off it.

However i'm entirely not sure this is correct... Could anyone explain how the Isotype Control Antibody helps the other antibody (eg.g CD 73) to bind more specific?

Explanation / Answer

I don't believe that the Isotype Control Antibody makes the antibody bind more specifically but rather ensures that your signal is specific to your antibody.

The Isotype Control Antibodies tend to bind to the non-binding domain of the antibody, typically the constant (Fc) domain. By measuring the concentration of the control antibody, you can determine what percentage of your light signal comes from your primary antibody and not from non-specific binders to your light source.

This is very similar to having secondary antibodies in your assay.

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