Plate layout & proper controls

This entry is part 1 of 1 in the series ELISA

In scientific experiments in general and in ELISA-style binding assay in particular, proper controls are of utmost importance. Without these an interpretation of the final result is often impossible. In planning a proper plate layout, you ensure that the signals you see, are really caused by your protein interaction and not by non-specific binding (NSB is a general problem in many interaction-assay we will later spend and whole article on.)

To detect non-specific binding, you need at least these controls:

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ELISA-style-binding assay

This entry is part 2 of 1 in the series ELISA

ELISA-style-binding-assays (here shortly called ELISA) are very robust and relatively simple established assays. They are extremely useful for making single points measurements, and thereby testing a lot of proteins for interactions. With a little bit more effort, it’s also possible to extract proper binding constants.

With a small series of posts, I’d like to help with the most important steps in planning and executing an ELISA assay.

Here the most important pro and cons of an ELISA assay in a table.

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ELISA simulator

In preparation of a series about setting up ELISA-style-binding-assays, I programmed a little tool, which helps to predict ELISA curves.

If you have an idea about the KD you can easily determine how many datapoints (samples) at which concentration you need to get a well defined curve. You can also easily compare the effects of different serial dilution regimes and different hill coefficients.  Hopefully this helps to better plan your next ELISA style binding assay!

You can find it in the menu under >Tools< ore directly here.

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A fresh start

Welcome to my little blog on BioAnalytics!

In this little blog I would like to share ideas, strategies and results around the general topic of protein-protein interactions and bioanalytical research on proteins. Some posts will be more directed to students or first-time-biochemists and these will be relatively instructional (like howtos and one side effect is that I can use them for my students). Other posts will be more directed towards experts and might be related to my actual research. I will try to publish something every Monday so stay tuned.

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