Antibody Specificity Profiling
Antibodies are essential tools of biomedical research. Immunoprecipitation, immunohistochemistry, immunofluorescence, ELISA assays, and the most common laboratory technique worldwide- the Western blot- all depend on them. As with computer programing, the adage “garbage in- garbage out” fully applies here. In fact, it has been speculated that the largest driver of poor reproducibility in published biomedical literature is antibody- driven. The rapid emergence of large-molecule therapeutics and early success in immuno-oncology (IO) are awakening the realization that these reagents must be characterized prior to use. In this sense, a slightly older adage has been turned on its head: Verify, don’t trust.
Array-based screening of mouse anti-POI* antibody (commercially obtained). A) Antibody to POI preferentially recognized fourteen other proteins among the >20,000 that were screened. B) Array image showing the region that contained TPRX1, the most preferred target of a-POI. C) Array image showing the region that contained POI (indicated by red arrow). The extent to which brightness and contrast required adjustment to even make POI visible highlights the distance between the intended target (POI) and preferred targets (like TPRX1). The green features in the upper left of the panel are controls for localization, but the features above and just slightly left of POI represent one of the fourteen other preferred proteins.
*In compliance with limited use liability we are unable to disclose the exact name of target or antibody manufacturer.
To get started with Antibody Specificity Profiling, contact us so we can discuss your specific project or idea!
Our Antibody Profiling Screen:
- Is far more informative than alternative techniques
- Is not affected by variable expression between or among samples (such as the one the manufacturer used to validate and the one you use in your lab)
- Operates against native or denatured proteins, depending on your intended use
- Eliminates epitope occlusion arising from multicomponent complex formation (native only)
- Is easily configured to capture PTM-specificity
- Provides information about epitopes of recognition and assists in evolving more specific reagents (see white paper)