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Question (76048) | BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

I have just bought the following item from your company : BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556).

I would like to normalize each well containing BrdU to the number of total live cells. What method would you advise me to use? I was thinking to normalize to the total number of protein or of DNA content but I can't see how these methods can be combined with the BrdU since the cells are immunostained and fixed.

If we understand your request correctly, you are trying to control to assure an equal number of viable cells is present in each assay well plus and minus BrdU. Our only suggestion would be for you to perform the assay in test tubes or deep well 96-well culture plates and count the viable cells prior to a short (˜2 hour) BrdU incubation. Alternatively, duplicate plates can be set up and one set can be assessed for cell number and one used for BrdU. Checking for protein or DNA content will not give an accurate cell viability count. Dead cells in the wells will have just as much protein and DNA as viable cells.