Although flow cytometry is a powerful technique for analysing the physical and chemical characteristics of cells, the appropriate choice and titration of antibodies are two crucial aspects. Why is this so? Quite simply because antibodies play an essential role in the detection and quantification of proteins on the cell surface or inside cells.
Optimal concentration
Each antibody has an optimal concentration at which it works best.
Too much antibody can saturate the binding sites, while too little may not detect the target effectively. Titration is used to find this optimal concentration, which is critical for reliable results.
Experimental reproducibility
Another key aspect is the reproducibility of experiments. Antibodies that are titrated consistently from one experiment to the next ensure that results are comparable. This is particularly important for longitudinal studies or validation between different studies.
Antibody costs
When titrated correctly, the amount of antibody used per experiment is very often less than the amount recommended by the manufacturer. Correct antibody titration will reduce the costs associated with the experiment.
In summary, antibody titration is an essential step in the flow cytometry process. It allows specificity, concentration and reproducibility to be optimized, ensuring accurate and reliable results.
Author: Philippe St-Onge, Core Facilities’ Echo committee
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The importance of titrating antibodies in flow cytometry
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The Core Facilities’ Echo