How to choose the right isotype control antibody for your experiments

Isotype control antibodies are antibodies that do not react with a specific antigen and are used as negative controls to correctly evaluate the results obtained from flow cytometry. immunohistochemistry and other applications.

In experiments using antibodies, background may occur due to non-specific binding to proteins other than the target or binding to Fc receptors on the cell surface. Therefore, in order to clarify that the reaction of the primary antibody to the target is specific and to accurately interpret the original signal strength after eliminating background effects, experiments using a negative control antibody are necessary. Since the binding of an antibody to an Fc receptor and the reactivity with a secondary antibody are specific to the antibody isotype, it is necessary to use the same isotype as the primary antibody against the target for the negative control. Here, we will introduce some points to consider when selecting and using the correct isotype control.

Choosing the right isotype control

Isotype control antibodies should be matched to the primary antibody against your target:

Host species and clonality
Class and subclass
Conjugation form

For example, if the primary antibody is a conjugated mouse monoclonal IgG1 antibody, then the isotype control should also be a conjugated mouse monoclonal IgG1antibody.

The concentration and reaction conditions for the isotype control should be the same as those for the primary antibody.

Abcam offers a wide range of unlabelled, Alexa Fluor® dyes , HRP, biotin, PE and FITC isotype control antibodies for various applications.