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How can knockout validation support the reproducibility of your experiments?
Dr. Alejandra Solache, Vice President of New Product Development at Abcam, recently discussed how we validate our antibodies using knockout (KO) technologies. Learn about the value of gene-knockout technologies in large-scale antibody screening by watching the video below.
Several studies have demonstrated the irreproducibility of life sciences reagents, which in the case of antibodies, frequently results from a lack of specificity to their targets of interest. This lack of specificity, related to issues such as cross-reactivity and off-target protein binding, has contributed to the so-called ‘reproducibility crisis’1,2.
KO validation is one of the most accepted and trusted validation processes for antibody specificity3,4. This robust technique can confirm the specificity of an antibody by testing it in a KO cell line, cell lysate or tissue that does not express the target protein. A specific antibody should, therefore, produce no signal in the KO cell line but give a specific signal in the wild type sample. In this way, KO validation serves as a true negative control5.
To ensure that our antibodies are specific and provide reproducible results, we apply KO validation at a large scale. We validate our antibodies in multiple applications, including flow cytometry, western blot, and immunocytochemistry, using multiple antibodies per target. Antibodies that demonstrate specificity are given a special “KO validated” designation, whereas those exhibiting off-target binding are removed from our catalog. We have now confirmed the specificity of more than 2500 antibodies with KO validation for over 2000 protein targets.
Dr. Alejandra Solache joined Abcam in December 2013 and manages Abcam’s Global New Product Development and validation teams in Cambridge, UK, Hangzhou, China as well as Eugene and Branford, CT in the USA. Alejandra has also been instrumental in developing Abcam’s innovation strategy and product pipeline.