siRNA knockdown validation 101: Incorporating negative controls in antibody research

F1000Res. 2016 Mar 9:5:308. doi: 10.12688/f1000research.8159.1. eCollection 2016.

Abstract

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.

Keywords: Western Blot; knockdown; negative control; shRNA; siRNA; specificity; transfection; vector.

Grants and funding

The author(s) declared that no grants were involved in supporting this work.