Isotype Control Flow Cytometry

Isotype control antibodies have no specificity for a target cell yet retain all the on specific characteristics of the antibodies used in an experiment.

Isotype control flow cytometry.

Isotype controls for flow cytometry and tissue staining isotype controls are important negative controls used to validate experimental results in flow cytometry and immunohistochemistry. Please the online available material and. 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. Considerations for the control of background fluorescence in clinical flow cytometry.

These antibodies having no antigen specificity should match the clonality isotype and conjugation of the experimental primary antibody. Flow cytometry isotype controls the use of isotype controls in flow cytometry is controversial and divides researchers herzenberg l et al. 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. Alternatively you can use our handy search table located at the bottom of the page to find the right isotype control for your experiment.

The purpose of such a control is to. Isotype controls are commonly used in flow cytometry experiments and immunohistochemistry ihc. Isotype control serves as negative control but the recent articles by flow cytometry society advocate that isotype control is not the right control. Confirm the specificity of primary antibody binding.

Applications for isotype controls. Selecting the appropriate isotype control may be an important element in flow cytometry experiments. Interpreting flow cytometry data. 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.

Maecker ht and trotter j. 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. 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.

The article also illustrates difference in undesirable binding at different levels using the same clone from different manufacturers.