Hello everyone,
I've done a few ELISA tests and I'm wondering about how to mix correctly my samples (cell culture supernatant) and the dilution buffer during an ELISA experiment.
For some experiments, I've directly mixed the samples and the dilution buffer in the wells of the ELISA plate, but carefuly, without touching the well walls and the bottom. I had good results at the end.
But my question is simple, is it ok to do that?
I know that it's better to do the mix before but with a lot of samples, It's easier to directly mix in the wells. About that, do you have any advices or tips on how to do the mix of samples and buffer, for exemple in plates, tubes?..
Thank you in advance
If you want to mix the sample and dilution buffer in the 96-well ELISA plate, you can avoid the risk of scraping the treated surface with pipet tips during mixing by using a plate mixer. The Eppendorf MixMate is a good one that has digital speed and time adjustments.
You could also premix the samples and dilution buffer in a 96-well polypropylene plate, then transfer the mixed samples to the ELISA plate using a multichannel pipettor.
Dear Thibault,
we always mix the supernatants directly in the ELISA plate with the dilution buffer. We never encountered any problems. Of course you have to consider, as Adam already mentioned, to pipette very carefully. But in general the ELISA plates are pretty robust even after coating with the capture antibody.
All the best and stay healthy,
Marc
Thank you for your answers,
About what you said @Adam B Shapiro, does the PP 96 plate has to be "low protein binding" ? Or any kind of PP 96 plate is good to mix samples before ?
Thank you again
Polypropylene plates are generally low protein binding, compared with polystyrene plates. I've never heard of low-protein-binding polypropylene plates.
You can also get deep-well 96-well polypropylene plates if you want to make solutions in one well to dispense into multiple wells.
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