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Positive control ELISA

elisa control

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3 replies to this topic

#1 julnobody

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Posted 26 January 2012 - 03:53 PM

Hi,

I performed some ELISAs recently that turned out positive (strong binding signals in most of wells, highly superior to no antigen control which showed signals close to 0).
However (shame), no positive controls were added to these plates...

Can I argue that positive controls would have been more useful to show that the assay had worked in case all my plates turned out negative, and that as I have strong signal in my sample wells it's not that much of a big deal not to have positive control added(except lacking rigor) ? ... To which extent can I analyze/interpret those results?

Thanks

#2 PAO_ahac

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Posted 27 January 2012 - 12:26 PM

did you run dose response curve? Did you run samples in duplicate?
the curve would contain negative...zero and vary concentrations of "postive"

Or, is this just samples and a negative?

#3 julnobody

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Posted 28 January 2012 - 02:21 AM

It's phage ELISA more precisely.(antigen coating, phage incubation, M13 Ab detection)

Samples weren't run in duplicates, no dilutions, it was just for checking the binding of my output phage.

A plate was run with +antigen and phage (except last row : +antigen and no phage),
Another one in parallel with the same phage samples but no antigen. (last row was antigen+no phage)

The +antigen plate was compared to the "background binding" plate..

#4 scolix

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Posted 30 January 2012 - 09:13 AM

always have a standard curve, if not possible, atleast have a positive and negative control for all experiments. These are going to tell if your experiment worked.





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