How to choose the correct antibody?

With millions of antibodies offered worldwide, by hundreds of suppliers, it may be a bit overwhelming, especially for beginners, to choose the correct antibody to answer a specific questions in a research project. Here is a short checklist with what to pay attention to:

Species reactivity – Has the antibody been used to detect the protein from the species we are working with? If not, a request should be sent to the supplier to confirm if the antibody may have so-called predicted reactivity to the target protein. If not, the antibody is not suitable, and false positive results can be produced.  

Validation data – Is validation data available on a product information sheet? For non-essential proteins, validation data should include results obtained with knockdown or knockout mutants. Western blot results showing that an antibody binds to a recombinant protein, are not relevant to the binding in a specific sample, extracted from a specific tissue or organism. Here we explain why.

Application – If the antibody has only been used in Western blot, it may not work in immunoprecipitation or immunolocalization. Each of these techniques requires separate validation data.

Recommended antibody dilution – The lower the dilution, the fewer tests can be conducted using an antibody. Here we explain why. 

Publication record – Being citet in scientific articles definitely confirms the performance of a given antibody. However, as there is no worldwide standard, product numbers are oftentimes not included in the "Material and Method" section. This makes it difficult to identify what antibody has been used to obtain the presented results. How to cite antibodies in scientific publication, is explained in this short video.