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standard for each plate?

standard for each plate? - ELISA Assay Forum

standard for each plate? - ELISA Forum. Discuss and post questions including optimization, troubleshooting, protocols and theory of ELISA assays using monoclonal, and polyclonal antibodies or peptides.


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Old 08-04-2009, 07:19 AM
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Default standard for each plate?



Hi All,
a very silly question may be-do we have to include the standards for each elisa plate we are doing or for eg, say if i am doing 3 plates at the same time then can i include the standards in just one plate,draw the standard curve and use the same values and equation for calculation of concentration of the other samples in the other plates.
hope anyone can understand what i mean to ask.
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Old 08-06-2009, 05:23 PM
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Default Re: standard for each plate?

Yes please use stds on each plate.
The reason is because the plates can have physical differences that can affect OD--this is a minimal concern.
But the main reason is because it can take large amounts of time between adding Ab or color reagent between plate 1 and plate 2 or 3.
Also human error can mean that one plate gets extra/less wash steps in between. Having the standard on each plate makes sure that this type of situation (and any effect from it) is minimized, since samples and stds are exposed to same variation(s).
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Old 08-06-2009, 05:29 PM
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Default Re: standard for each plate?

As a further note: I've actually recently did this same thing, ELISA on three plates.
While the std curves were nearly identical and there was a tight CV from sample duplicates on all plates, the fact is there is enough slight difference, that using only one std curve for all three plates would have resulted in enough rounding error to compromise downstream (i.e. statistical) analyses.
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