If cells obtained from different passages, with different plating but harvested on the same day are used in a single plate to take an ELISA measurement (with the given standards and treated plate), is this still a biological replicate? Or is it different? Please provide me with some clarifications.
Thanks in advance!
I would recommend a replication with another batch of experiments, i.e. harvest and generate data on monday/tuesday, then replicate the same thing thursday/friday. Passaging the cells doesn't make it a biological replicate, they resemble technical replicates as they are the same clone. It is crucial that two different experiments give similar results to ensure there is reproducibility. I have seen results that seem exciting, we replicated it and wasn't able to reproduce it. Don't make claims based on a single experiment in a single day, it is assuring that a different set of experiments on a different day can tell a consistent story that align with your hypothesis and initial findings. The means and P-values will be different, but hopefully the vector of the change is similar. At least 1 replication on a different day would be compelling evidence.
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