Have you heard of The Domino Effect? I’m sure you have but let me remind you what it is. The Domino Effect is the cumulative, often disastrous effect produced when one single event sets off a chain of other events, the last one depending on the very first one.
In the context of your daily lab activity, The Domino Effect is at play when you discover the disastrous results you might get when you don’t put enough time and effort into planning your experiments from the very first step. Read on to find out how you can prevent this catastrophic chain of reactions when it comes to antibody-based experiments.
Whether you are a beginner or a veteran researcher, you have surely experienced, heard stories, or know someone who has cautionary tales to tell about experimental failures. Planning an experiment is a delicate matter and genuine mistakes can occur at every level. However, although you must worry about every single step during your planning process, as each of them represents a crucial juncture, it is true that the first step can set the direction of the whole experiment.
One target, different results?
The most common and easy mistake that happens in the lab is due to poor attention when choosing and preparing reagents. Buffers and chemicals are all good candidates. But think about the majority of biological assays run daily in the lab: immunohistochemistry, immunocytochemistry, western blot, enzyme-linked immunoassay, flow cytometry, and immunoprecipitation. What do they have in common? Antibodies! They all rely on a specific antibody-epitope binding. These antibodies may be monoclonal, polyclonal, or recombinant from different organisms, and they may be used to interrogate biological systems and signaling pathways on normal, cancerous, or other tissues.
Choosing a highly validated and well-characterized antibody is a fundamental step and is certainly an essential starting point to drive your results in the desired direction. It is not about good and bad antibodies, but antibodies that can be trusted, that are reproducible, and that work in specific tissue-application combinations. Even a good antibody can fail if used in the wrong context.
So, if you do have a choice, look for an antibody you can trust and increase your chances of success. But how can you trust an antibody?
The dictionary definition of trust is “the belief that someone or something is being truthful”. Trust is a mixture of character and competence, in other words, your strengths, what you do, and the results you produce.
The character and competence of an antibody are defined, at least, by three characteristics such as specificity, selectivity, and reproducibility – in relation to the context in which it will be used. Although these are all equally important features, they depend on different concepts.