Isotype Control Flow Cytometry Gating

Gating out dead cells using live dead marker isoclonic controls.

Isotype control flow cytometry gating. This is where cells are stained in the presence of an excess of identical unlabeled antibody. Clinical cytometry 76 6 355 364. The various positive controls are used for compensating and gating when setting up the flow cytometer. They should not be used to distinguish positive from negative cells or set positive gating regions.

Recommended controls for flow cytometry facs along with the samples to be labeled the following controls should be used whenever possible. 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 isotype control becomes another variable to be tested validated and optimized for marginal gain as a gating control. The section of the above paper focusing on isotype controls summarizes the problems with their use very clearly.

Considerations for the control of background fluorescence in clinical flow cytometry. Control sample type primary ab secondary reason cells only use treated and untreated cells. The article also illustrates difference in undesirable binding at different levels using the same clone from different manufacturers. Spurgeon and khalid m.

The unlabeled antibody takes up all the binding sites preventing the labeled antibody from binding specifically. An ideal isotype control should. Considerations for the control of background fluorescence in clinical flow cytometry. As maecker and trotter state it is thus a hit or miss prospect to find an isotype control that truly matches the background staining of a particular test antibody.

Instrument setup controls and panel performance benjamin e j.