Isotype Control Flow Cytometry

You may choose not to use isotype controls but if you do there are some simple rules to follow which will ensure you use them properly and in conjunction with other controls.

Isotype control flow cytometry. Selecting the appropriate isotype control may be an important element in flow cytometry experiments. Isotype control serves as negative control but the recent articles by flow cytometry society advocate that isotype control is not the right control. Isotype controls are commonly used in flow cytometry experiments and immunohistochemistry ihc. Considerations for the control of background fluorescence in clinical flow cytometry.

Flow cytometry isotype controls the use of isotype controls in flow cytometry is controversial and divides researchers herzenberg l et al. Isotype controls for flow cytometry and tissue staining isotype controls are important negative controls used to validate experimental results in flow cytometry and immunohistochemistry. Alternatively you can use our handy search table located at the bottom of the page to find the right isotype control for your experiment. Applications for isotype controls.

Although isotype controls are mainly used in flow cytometry they can be used as standard blocking agents and protein coating agents for other applications including immunofluorescence immunocytochemistry western blotting and elisa. Please the online available material and. Confirm the specificity of primary antibody binding. For these applications and other immunoassays including elisas western blotting and immunocytochemistry icc an isotype control antibody can also serve as a blocking or coating agent.

Interpreting flow cytometry data. The purpose of such a control is to. Isotype controls should be used to determine the background due to nonspecific antibody binding. Maecker ht and trotter j.

An isotype control uses an antibody of the same isotype as the primary antibody but is specific for an antigen absent from the cells under study. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. The article also illustrates difference in undesirable binding at different levels using the same clone from different manufacturers. The section of the above paper focusing on isotype controls summarizes the problems with their use very clearly.

To see our range of flow cytometry isotype controls with information on how and when to use them download our isotype controls brochure.