Antibody validation emphasised in the new version of the Human Protein Atlas

Figure 1. Validation with independent antibodies.
Figure 2. Validation with orthogonal methods.
Figure 3. Validation with capture Mass Spectrometry.

Research-used antibodies continue to be important tools for detection, analysis and characterization of proteins. As reagents, and used in combination with technological platforms, antibodies enable researchers to explore the human body and understand how building-blocks like proteins are produced, distributed and diminished, as well as monitor alteration in time, space and during disease development.

Although many antibodies are available the uniqueness and the performance of these reagents is constantly questioned and remains one of the major challenges in proteomics research.

Since the initiation of the Human Protein Atlas project development of antibody as reagents and testing their usefulness has been a major undertaking.

A new version of the database published on December 1, 2017 contains the latest achievements regarding enhanced validation of antibodies. The enhanced validation has been defined based on the recommendations from the International Working Group for Antibody Validation (IWGAV) published in Nature Methods in 2016 as an international collaboration representing the research community involved in antibody research (Uhlen et al. 2016).

In version 18 of the Human Protein Atlas 10,540 antibodies targeting a total of 6,787 human proteins have enhanced validation data. Descriptions of the five validation strategies are available on the Human Protein Atlas and outlined in the press release.

As an example the antibody HPA023871 targeting the LIM domain and actin binding 1 protein was validated using three of the five strategies, namely independent antibody validation (Figure 1), orthogonal validation (Figure 2) and capture Mass Spectrometry validation (Figure 3).

The enhanced validation is presented on Human Protein Atlas portal for every protein target in the "Antibody validation" section. The detailed information regarding validation of each antibody and images from experimental results are displayed together with a summary.

This resource can be used to scrutinize and assess performance of the antibodies and uniqueness of the recognized epitope.


Uhlen et al 2016, A proposal for validation of antibodies, Nature Methods 13, 823-827, DOI:10.1038/nmeth.3995.

Cristina Al-Khalili Szigyarto