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Antibody Verification |
Antibodies are broadly used proteomics tools that play a crucial role in various research disciplines. Poor antibody validation and application performance often result in inconsistent data and reproducibility of results. Inadequate antibody performance and lack of specificity poses significant risk to researchers causing erroneous data, poor data quality, wasted time and resources.
Our commitment to antibody advanced verification
To drive reagent quality, directly address antibody reproducibility, and help provide customers with confidence in our antibodies, Thermo Fisher Scientific has adopted three concepts to help improve experimental reproducibility and reporting based on the recommendations of the International Working Group for Antibody Validation (IWGAV):
| IWGAV Recommendation | Thermo Fisher Scientific’s Antibody Verification Effort |
|---|---|
| Expanded specificity testing methods to ensure superior antibody results | At Thermo Fisher Scientific, we have continued to expand our testing methods to cover more and more of our antibody portfolio. Details on our advanced verification and application verification methods will be reviewed below. |
| Increased visibility of verification data so researchers can make a better antibody selection | All antibodies on the Thermo Fisher Scientific website that have undergone and passed the advanced verification testing are identified with an “Advanced Verification” badge. The badge can be found in the antibody search results and at the top of the product detail pages. See figure 1 for an example of the advanced verification badge. Data supporting the antibody’s advanced verification status can be found in the data galleries on the product pages as well. |
| Improved reporting of antibody specifics such as clone, isotype identity, and functional characteristics to facilitate the replication of published results | Thermo Fisher Scientific reports all antibody specifics on the individual product detail pages. |
Figure 1. Example of what an antibody that has undergone Advanced Verification looks like in the search results. The red check and Advanced Verification tag will be present here and on the top of the product description page.
At Thermo Fisher Scientific, we help provide confidence in selecting the right antibody for the intended research needs to enable researchers to successfully reproduce and confirm experimental results. This means that an antibody must be specific and selective within the context it is being used. To achieve this, we use a comprehensive approach to antibody verification that is tailored to the antibody target and the relevant application. Our advanced verification occurs in addition to standard antibody testing that happens during manufacturing.
Advanced verification
Advanced verification is additional testing that verifies that an antibody will bind to the correct target. We consider two key factors when selecting the appropriate advanced verification testing method(s):
- The nature of the target—this includes considering relevant target properties such as its biological function, sub-cellular localization, post-translational modifications, gene essentiality, and the rate of turnover.
- The nature of the sample type—the sample type can influence epitope exposure and the reactivity of an antibody in a specific application.
Click on each testing method below for detailed testing strategies, workflow examples, and example data.
Expression testing using either CRISPR-Cas9 cell models or RNAi to knockdown genes of interest.
Measurement of target expression is performed using two differentially raised antibodies recognizing the same target.
Detecting downstream events following cell treatment.
Using naturally occurring variable expression to confirm specificity.
Functional blocking of protein activity by antibody binding.
Using arrays to test reactivity against known protein modification.
Using SNAP-ChIP to test reactivity against known protein modifications.
Testing using immunoprecipitation followed by mass spectrometry to identify antibody targets.
Application verification
We also help ensure that the antibody works in particular application(s) of interest. Application verification is used to help ensure that the antibody works in a singular application, as well as in multiple other applications to demonstrate that the antibody binds the correct target. We cover a wide range of applications including western blot, flow cytometry, ChIP assays, immunocytochemistry (ICC), and immunohistochemistry (IHC). Click on each of the applications below to see a typical protocol that we use for application verification.
The history of antibody verification at Thermo Fisher Scientific
Antibody validation through advanced specificity verification
Addressing the antibody reproducibility crisis: A panel discussion with key scientific leaders
*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.
For Research Use Only. Not for use in diagnostic procedures.
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