I am under training and just running controls
positive 1 & 2 , negative control and titer controls. My controls are higher and hence my plates are failing. Can you think of possibilities as to why my controls fall in higher range?
It is very simple, If you are getting high signals then you just dilute your samples to get O.D of low intensity which will in your range of ELISA plate. OK
Try doing a checker board titration to determine the appropriate dilution factor to use and voila you will get your absorbances right.
I agree with the comments above. Do a serial dilution and see which dilution fits your samples between the positive & negative controls. :)
As the above comments say, dilute your samples. If you're using an ELISA kit, there should be either a diluent for you to dilute with, or the instructions should contain a recipe (0.1% or 1% BSA in PBS is fairly common). An alternative might be cell culture medium, if these samples are from cells. When you're running the samples, you will probably end up with a good idea of how much you need to dilute to get all results within the standard curve, but it's sometimes a good idea to run samples twice, at two dilutions, if you have space on a plate, and especially if you don't want to freeze-thaw your samples too much. Hope this helps!
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