Mastering Post-Translational Modifications

 

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).

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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.


Featured Products

Product Catalog #
Histone H3 K27 Methyl Antibody 200-401-M74
Histone H3 K27ac Antibody 600-401-K00
Histone H3 K27me3 Antibody 600-401-J99
Histone H3 K4me2/pT6 Antibody 600-401-K01
Histone H3 K79Me3 Antibody 600-401-I95
Interferon-alpha Receptor Type I phospho S535/ phospho S539 Antibody 600-401-D67
MeCP2 phospho S421 Antibody 600-401-X14
MeCP2 phospho S80 Antibody 100-401-D71
Histone H3K23me2 Antibody 600-401-K02
NMDA NR2B Subnit phospho S1166 Antibody 600-401-X17