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All tags Jane Women's Snapper Flat Mary Sterling Keen City SH7qIIAPrimary antibodies Knockout (KO) validated antibodies: specificity confirmed

Knockout (KO) validated antibodies: specificity confirmed

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With the increasing need for highly specific antibodies, Abcam introduces KO validation as a standard quality control protocol for antibodies at scale


The need

Affinity binders/antibodies are amongst the most commonly used tools in life science to study proteins and their functions within various biological pathways and diseases. With their specificity and sensitivity, these tools can help to easily identify the protein of interest at various expression levels. However, an increasing number of studies have shown that not all of these tools are specific, leading to a growing issue of experimental irreproducibility. This irreproducibility is leading to wasted time, resources and, unfortunately, the retraction of papers within the scientific community (Bradbury & Plückthun, 2015; Baker, 2015; Mullane et al., 2015Women's Calf Buckle Boot Brian Detail Boot Souls Gentle with Black Angled Mid Leather Topline Harness 5TSqxISgn1).

The solution: knockout validation

One of the most accepted and trusted validation processes for antibody specificity is through knockout validation (Madhusoodanan, 2014; Rhodes & Trimmers, 2008). In a knockout validation, antibody specificity is confirmed by testing the antibody of interest on a knockout sample or cell line which doesn't express the target protein.  In a knockout cell line, the antibody target is not present as the gene encoding the protein of interest is eliminated or "knocked out". This data is compared side-by-side against a normal cell line (or wild type) and if the antibody is truly specific, the antibody should have no detection in the knockout cell line, but instead a specific signal in the normal cell line. Knockout validations offers a true negative control to confirm the antibody specificity to the protein of interest (Bordeaux, et al., 2010).

Knockout validation as a part of Abcam's standard antibody validationAnthracite Marylin 03 Marylin 03 wPg0q

Abcam has incorporated knockout based validation as a standard level of validation for its antibody products. We are starting at 500 antibodies a year, and we will knockout validate these antibodies as a part of our rigorous antibody validation process.

When you see the knockout validated seal, you can trust that the antibody has not only been validated in the recommended applications and species, but its specificity has been confirmed through our in-house knockout validation approach.

Abcam knockout validates its antibodies using the extensive library of knockout cell lines available from Horizon Discovery.


  • See how KO validation compares to other types of antibody validation assays

  • Learn more about the technology behind our KO cell lines

  • See different scenarios of results from KO validation 
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See which antibodies have been knockout validated


We want to hear from you

If you have any comments or questions, please feel free to contact us by phone or email, or by simply filling out an online inquiry form.




​References​​​​​​​

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  • Bradbury A, Plückthun A (2015). Reproducibility: Standardize antibodies used in research. Nature 518(7537):27-9. doi: 10.1038/518027a.
  • Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature 521(7552):274-6. doi: 10.1038/521274a.
  • Mullane K, et al., (2015). Guidelines for manuscript submission in the peer-reviewed pharmacological literature. Biochem Pharmacol. pii: S0006-2952(15)00358-5. doi: 10.1016/j.bcp.2015.06.023.
  • Madhusoodanan J (2014) .  Validating Antibodies: An Urgent Need.  The Scientist.
  • Rhodes KJ, Trimmer JS (2008). Antibody-based validation of CNS ion channel drug targets. J Gen Physiol. 131(5):407-13. doi: 10.1085/jgp.200709926.
  • Bordeaux, et al., (2010).​ Antibody validation. Biotechniques 48(3):197-209. doi: 10.2144/000113382.​​​


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