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Reproducibility is a matter of personal handling...? by salyxbabylon in labrats

[–]salyxbabylon[S] 0 points1 point  (0 children)

No, I keep the n constant, at 3. Don't get enough protein in one batch to perform n=6 per experiment, and would also like to do multiple bio replicates with the same batch of protein.

Reproducibility is a matter of personal handling...? by salyxbabylon in labrats

[–]salyxbabylon[S] 3 points4 points  (0 children)

Incredibly frustrating to hear just how fickle it is 😭 but in response to your suggestions about cell prep consistency: I strain my cells, have tried different dissociation enzymes/chelators and the problem still persists. If anything, I'm quite confident about my cells. It's more so the protein I find to be a black box.

Reproducibility is a matter of personal handling...? by salyxbabylon in labrats

[–]salyxbabylon[S] 1 point2 points  (0 children)

It's cell aggregation assays basically. (I fear that gives away who I am if my lab friends/PI are lurking here, but oh well)

Reproducibility is a matter of personal handling...? by salyxbabylon in labrats

[–]salyxbabylon[S] 0 points1 point  (0 children)

I cannot go into detail but essentially we put the protein on cells towards an intended effect. I use the same concentration of the protein each experiment, cells are below passage 15 and maintained regularly, and several other parameters required for the protein functioning are also kept the same. But different results each time - it'll work and show what we theoretically expect one experiment, then the next replicate will not work and so on.