Post-translational modifications (PTMs) play a key role in dynamic cellular processes, regulating gene expression, protein activity, localization, and degradation, as well as protein interaction. Modification-specific antibodies offer a versatile tool for the characterization of post-translational modifications. Learn how you can choose the best high-affinity, high-specificity antibody for your PTM detection needs.
At Rockland, scientists have developed proprietary methods for the development of highly specific PTM antibodies that can be used in a wide range of in vitro and in vivo studies of a modified protein, some of which are not easily performed by other approaches, such as mass spectrometry (MS).
Antibody Selection Tips for PTMs
1 Preparation
From an antibody production point of view, the differences between modified proteins can be quite small. Peptide design and immunogen quality are critical to the generation of a specific immune response to ensure the production of high-quality antibodies.
2 Production
Antibodies against PTMs are generated using a short, specific region of the protein, largely eliminating the issue of specificity seen with antibodies generated using large constructs as immunogens. However, it is critical that the antibody be tested against established positive and negative controls to ensure specificity for the modification. Polyclonal antibodies can be immunodepleted during production if the sample contains antibodies that recognize other PTMs.
3 Validation
Dot blot assays and ELISAs can be used to assess both antibody specificity and sensitivity. Keep in mind that, in addition to being specific for the required modification, the antibody must be validated for the application of choice using appropriate positive and negative controls.