Isotype Control Flow Cytometry Definition

Isotype controls should be used to determine the background due to nonspecific antibody binding.

Isotype control flow cytometry definition. To see our range of flow cytometry isotype controls with information on how and when to use them download our isotype controls brochure. The purpose of such a control is to. 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. Confirm the specificity of primary antibody binding.

Flow cytometry uses fluorescently labeled molecules to identify and characterize antigen profiles in a variety of research applications and for clinical diagnostics on a single cell basis. Flow cytometry isotype controls the use of isotype controls in flow cytometry is controversial and divides researchers herzenberg l et al. 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. The article also illustrates difference in undesirable binding at different levels using the same clone from different manufacturers.

The host species ig subclass iga igg igd ige or igm of the primary or secondary antibody. Maecker ht and trotter j. Stem and progenitor cells by flow cytometry time to let go. The isotype control should match.

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 antibodies have no specificity for a target cell yet retain all the on specific characteristics of the antibodies used in an experiment. Applications for isotype controls. The section of the above paper focusing on isotype controls summarizes the problems with their use very clearly.

Isotype control cells incubated with isotype control antibodies antibodies usually raised against an antigen that should not be present in your cells. Isotype controls are used to determine the background caused by nonspecific antibody binding. To determine nonspecific binding of the primary antibody. Herzenberg l tung j moore wa herzenberg la and parks dr.

Isotype controls in the analysis of lymphocytes and cd34. Alternatively you can use our handy search table located at the bottom of the page to find the right isotype control for your experiment. Selecting the appropriate isotype control may be an important element in flow cytometry experiments. Our mouse isotype reagents are based on monoclonal mouse antibodies of different isotypes and unless indicated otherwise directed towards aspergillus niger glucose oxidase an enzyme which is neither present nor inducible in mammalian tissues.