Antibody Identity card
Introduction
The selection of the appropriate antibody (Ab) for your research is currently challenging due to the high number of Abs sold by more than 300 companies that are accompanied by limited correct validation and characterization data. This makes it extremely difficult for the scientific community to choose the right Ab for their particular research area.
With this in mind, we have launched the Ab ID CARD initiative that is aimed to highlight the fact that every Ab has its own specific identity and it is essential to report the properties and performance of each individual antibody in its laboratory applications in a transparent and understandable way. The motivation behind this initiative is to share our practical experience of Ab validation and characterisation with other members of the research community (MAbs 2016, 27-36 2015, doi: 10.1080/19420862.2015.1100787 - PDF 1.1 MB).
Diagram of the Ab ID card
The first part of the Ab ID CARD shows information on the protein of interest as well as some very useful links that will provide an overview of your target molecule (e.g. protein sequence, existence of variants/isoforms, transcript expression from microarray data, protein expression data etc.).
This is followed by a list of reported Abs against your target. These Abs are been identified by their clone name and willhave been tested in several applications following the EuroMAbNet Ab validation guidelines.
Clicking on each Ab clone name shows all the information available for that Ab of interest (e.g. immunogen used, epitope if known, etc.) as well as the protocol and concentrations used for each application. Our approach was to test several Abs against the same target produced using different antigens and species in parallel since observing any common patterns can significantly strengthen the confidence in the validation data generated. We have also studied cross-species reactivity for some antibodies.
Clicking on the boxes shows the results obtained in the various different applications (e.g. WB, IHC etc) for the Ab. A colour code (green - working, orange - no conclusive, red - not working, grey - not tested) indicates Ab performance in each application.
This information allows you to try to reproduce our results in your own laboratory. Please note that researchers perform similar assays in many different ways and so we cannot guarantee that the results obtained during our validation process may differ using different laboratory conditions.
The creation of this platform, providing access to information about different Abs and how they perform across techniques, strengthens our commitment to increase research traceability and reproducibility. We know that this project is ‘a drop in the ocean’ but we believe that this and other small steps done by both the research community and industry will finally reduce frustration in researchers’ time and effort and improve the advance of reproducible science.
We would like to thank the Confocal Microscopy (Manu and Jesus), Immunohistochemistry (Patri and Zaira), Cytogenetics (Raul and Sandra) and Flow Cytometry Units (Lola and Julia) of the CNIO Biotechnology Programme for their outstanding work done in the validation and characterisation of the mAbs reported in the Ab ID Card.
We would also like to extend our deepest gratitude to Marcos de Miguel and Dr Karen Pulford for the design and writing of the Ab ID CARD web.