Poor quality, unverified antibodies are a major factor underlying the crisis of reproducibility in research, resulting in an estimated $800 million of waste each year and a ten-fold spike in retractions over the past decade.
More antibody validation protocols would identify non-specific reagents – a major source of irreproducible research – with the inclusion of negative controls. This article presents an overview of one such method: siRNA knockdown. The authors outline a general protocol, the knockdown mechanism, and tips for evaluating knockdown experiments.