Hi,
I have run 16-plex and for some markers and in some plates the standard curve was not clear for the last standard dilution, but in other plates all points worked. I'm interested in calculated the fold change in the markers after vaccination, then measure all values is important for me. It would be ok if I use the standard curve where all point worked in another plate? or if I pool for the last standard dilution from different plates (where it worked) and use it in the plates that didn't worked.
All the plates have the same standard lote and were run in the same on after each other.
/Paola
When you say that last point isn't clear... is it because it's below the blank?
So... technically, you are suppose to use the standard curve that is plated on the plate. Because of plate to plate variations.
However, having said that.. it is acceptable to combine standard curves... Now... there is a criteria that is usually needed for this...which are control samples from plate to plate. These control samples have known concentrations and are designed to fall onto specific points on the standard curve. So that when standard curves are averaged, these controls should still fall within a QC cutoff.
But... for your purposes, it should be acceptable to average all the standard curves from all your plates and use it for analyses across all plates. In your publication, you just have to detail what you did.
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