Dear Albert, Sorry, but I don't agree with you. When you should look at a marker 80% of your cells is positive for. And you determine the amount of antibody (= your concentration) for 300.000 cells. It is not possible to get even a peak far from your negative peak when you stain 5 million cells. This means that you throw away at least 5 times the amount of antibody when you stain 300.000 cells. Because you determined (titrated) your antibody for 5 million cells. And the definition of concentration is the amount of Ab in a certain volume. So when you got 100.000 cells and need 0.01 µg of Ab to stain them you can add this in 50 µl or in 5 ml. The only difference is the timeperiod it will take to get your Ab on the cells. Or am I mistaken? Best regards, Joost Schuitemaker
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