A commitment to antibody performance

Every year, researchers waste valuable time and money using antibodies that do not perform as intended. Towards that end, Thermo Fisher Scientific is committed to adopting validation* standards for our Invitrogen antibody portfolio in line with the recommendations of the International Working Group for Antibody Validation (IWGAV).

Using our new standard, Invitrogen antibodies are being tested for both target specificity as well as application validation* to ensure proper functionality in researcher’s experiments. By doing our part in the rigorous testing of our antibodies, Thermo Fisher Scientific will help researchers efficiently save time and money using antibodies that work for their research needs.

Specificity verification

Which validation pillar would you like to explore?

Immunoprecipitation-mass spectromery (IP-MS) Genetic modification Independent antibody verification (IAV) Orthogonal (functional) testing
IP-MS—testing using immunoprecipitation followed by mass spectrometry to identify antibody targets

Knockout—expression testing using CRISPR-Cas9 cell models

Knockdown—expression testing using RNAi to knockdown gene of interest

Endogenous control—using naturally occurring variable expression to confirm specificity

IAV—expression of target protein using two antibodies with non-overlapping epitopes

Cell treatment—detecting downstream events following cell treatment

Neutralization—functional blocking of protein activity by antibody binding

Peptide array—using arrays to test reactivity against known protein modifications


Resources on antibody verification

Learn more about efforts from the research community to develop and implement standards for antibody validation.

IWGAV paper

Workshop report and session recordings

Webinar


Application validation

Most antibodies were developed with specific applications in mind. For example, when developing a ZO-1 antibody, a researcher’s initial interest and priority may be to obtain an antibody that provides excellent tight junction staining in immunofluorescent (IF) imaging. Subsequently, the researcher may want to perform a western blot, tissue staining, ELISA, or some other application with the same antibody. Only knowing that an antibody is verified for a specific target does not mean it will work well in every application. Also, an antibody that is optimal for one application may not be the very best for another application.

Testing that an antibody generates acceptable results in a specific application is the only way to confirm antibody performance. There are a tremendous number of variables, besides the antibody, that can impact a functional testing experiment. From upstream factors like sample type and quality of preparation, to downstream factors like detection methods and instruments, multiple variables should be considered.

Our commitment to functional testing

We are currently retesting the portfolio of Invitrogen antibodies to validate them in different applications. The antibody testing team consults with experienced biologists to select and plan the tests, being sure to include the samples types and treatments most relevant to a given target.

Our functional tests include these applications:

  • Western blot
  • Immunofluorescent (IF) imaging
  • Flow cytometry
  • ChIP
  • Immunohistochemistry (IHC)

Test results will be reported in many places throughout the Invitrogen antibody portfolio:

  • On the product pages: Data images will be presented with extensive figure legends detailing how the data was generated, including any special tips relating to sample prep or data collection. The product pages will include both applications tested in-house and applications that have been published, including the dilutions used in the publication and a link to the publication.
  • On the product: Suggested antibody dilutions will be provided when possible.
  • On the antibody search tool: Search filters will allow scientists to quickly identify applications that have been shown to work in functional testing for each antibody.
Data provided for NF-kB p65 antibody after retesting

The data provided for NF-kB p65 antibody (Cat. No. PA1-186) after retesting. Prior to retesting, our PA1-186 antibody only had western blot data provided (top left). After retesting we were able to add immunofluorescent microscopy, flow cytometry, and chromatin immunoprecipitation (ChIP) data on the product page, including suggested dilutions for each of the applications.


*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.