Hello,
I am planning to run a series of ELISA experiments and am asking for basic tips for improving the quality and reproducibility of my experiments. I have only done the experiment once, but my results were variable.
I would like this thread to be helpful to not only myself but other learning this technique as well. That being said, if anyone has small things that they do that would not be explicitly explained in an ELISA protocol what are they? Please explain one's rationale.
Thanks everyone.
The number of factors are affecting the results of ELISA. If you take care of all surely you are able to get excellent result.
First and most important is to standardize you technique. Confirm the minimum concentration of your test biomolecule giving results repetitively. Fix the concentrations of antigens and antibodies being used in ELISA. Fix your negative and positive control which will again add to good quality of results. Take care of incubation temperature, calibrated pippets, temperature of reagents being used in ELISA are all key steps to improve the quality of results. Run each sample in triplicate so as to confirm the intervariation which will give you an idea of probable source of error. Surely quality of sample again decide the results. So many factors to be handled carefully to get best possible result.
With very best wishes for next experiment.........
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