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Incubation conditions?!? - Gain control over low ODs (Oct/18/2008 )

Hello to you all!

I'm rather a newby on Elisas and I am still struggling to get control over my ODs. They are really low, even for the defined standards. As I'm sure to use the right/recommended solutions or have checked on different options I now fear the conditions under which I perform the Elisa are not optimal.
Can anyone tell me incubation conditions (e.g. temprature, time,...), that made Elisas work properly in your lab?

Thank you in advance for your Help!


If this is a purchased kit follow the directions.

I would suggest finding a commercial kit that mimicks your system and follow their incubation times (and temperatures) as a starting point.

Incubation times and temperatures vary from assay to assay...times can be as short as 15 minutes to several hours. Temperature usually ambient or 37C.


Thank you for your answer.

It is/are purchased kits. I get the same kind of "error" with different assays. So I fear that I'm making a systemic mistake, but I cannot find out, what it is.
Not having done an ELISA before I made really sure to follow the instructions by the word. I now hope that a variation of the condition could lead to better signals!

Kind Regards


Besides the incubation conditions...if the conjugate is in concentrate form do a few points using 2X more concentrated conjugate. Your signal will increase...hopefully the background will remain constant.

Also, if you are using diluted samples use them in more concentrated....this will also boost your signals.

Since it is a commercial should definitely call the manufacturer for assistance.