How Are Your Antibodies Validated?

Stop wasting time and money on unvalidated antibodies

In September 2015, Thermo Fisher Scientific helped organize and supported a group of leading researchers from institutions throughout the world to draft a set of proposed standards for the best ways to validate* antibody specificity. The International Working Group for Antibody Validation (IWGAV) published their proposed strategies in Nature Methods in September 2016.

In the fall of 2016, many of the IWGAV members, as well as representatives from antibody producers, pharmaceutical companies, and leading academic research labs, met at an event coordinated by the Global Biological Standards Institute (GBSI) to further study this topic. This meeting reviewed and validated the IWGAV’s proposed standards and discussed ways to implement the standards across the industry.

As a result of our active discussions and key participation in these forums, we have seen a clear consensus develop around several impactful practices to help further advance antibody validation*. These new guidelines will add to the extensive application validation* already performed for Invitrogen antibodies and will focus on confirming antibody specificity. Testing antibodies for application validation* is critical and must be carried out with the utmost rigor, but it can be a further challenge to confirm antibody specificity. By combining both application validation* and specificity testing, we aim to ensure that antibodies meet the highest possible standards of overall antibody validation.*

These specificity verification methods are being used to systematically validate* the Invitrogen antibody portfolio.

The Invitrogen antibody portfolio

Invitrogen antibodies have more than 120,000 supporting images and more than 60,000 reference citations. Stay tuned as we continue to establish new standards in antibody testing and verification.

Advanced verification testing methods

Immunoprecipitation-mass spectrometry


Immunoprecipitation-mass spectrometry (IP-MS)
: testing using immunoprecipitation followed by mass spectrometry to identify antibody targets

Genetic modification
  • Knockout: expression testing using CRISPR-Cas9 cell models
  • Knockdown: expression testing using RNAi to knock down gene of interest
IAV


Independent antibody verification (IAV)
: measurement of target expression is performed using two differentially raised antibodies recognizing the same protein target

Biological verification
  • Cell treatment: detecting downstream events following cell treatment
  • Relative expression: using naturally occurring variable expression to confirm specificity
  • Neutralization: functional blocking of protein activity by antibody binding
  • Peptide array: using arrays to test reactivity against known protein modifications
  • Orthogonal method: correlation using both antibody dependent and independent method
Immunoprecipitation-mass spectrometry


Immunoprecipitation-mass spectrometry (IP-MS)
: testing using immunoprecipitation followed by mass spectrometry to identify antibody targets

Genetic modification
  • Knockout: expression testing using CRISPR-Cas9 cell models
  • Knockdown: expression testing using RNAi to knock down gene of interest
IAV


Independent antibody verification (IAV)
: measurement of target expression is performed using two differentially raised antibodies recognizing the same protein target

Biological verification
  • Cell treatment: detecting downstream events following cell treatment
  • Relative expression: using naturally occurring variable expression to confirm specificity
  • Neutralization: functional blocking of protein activity by antibody binding
  • Peptide array: using arrays to test reactivity against known protein modifications
  • Orthogonal method: correlation using both antibody dependent and independent method

To find out more, go to thermofisher.com/antibodyvalidation

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Over 74,000 high-quality Invitrogen antibodies, covering 85% of the proteome

* The use or any variation of the word “validation” refers only to research use antibodies that were subject to functional testing to confirm that the antibody can be used with the research techniques indicated. It does not ensure that the product(s) was validated for clinical or diagnostic uses.

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For Research Use Only. Not for use in diagnostic procedures.