Alternatives to Phospho-specific Antibody Assays

We at Novus Biologicals have a broad antibody database of both standard and phospho-specific antibodies, i.e. those targeting only the phosphorylated component of a protein. Since phosphorylation is switched on and off depending on the protein’s active state, these are immensely useful when investigating cellular pathways and diseases.

As well as antibodies we also supply proteins and peptides for research. Antibodies are not always required during phosphorylation studies – for example, phospho-proteins play a role in proteomics, the study of all the genes in a genome.

Phosphorylation can be studied directly using ion traps, which detect the mass difference that signals the event. Electron transfer dissociation, which can fragment and sequence peptide ions is an advancement of this. Unlike the more common collision-induced dissociation technique, the bond between the phosphate and its attachment point remain intact, so scientists can sequence the peptide backbone and determine where the phosphate originated. It is of particular use in post-translational modification studies.

Ion traps are commonly used for initial phospho-detection work. Gel staining using a mass spectrometer is another method. Whether you use ion traps, antibodies or a spectrometer for your phosphorylation studies, it’s important your samples are well prepared beforehand. It’s particularly important you use high-level purification techniques and, in the case of antibodies, use only the purest and highest quality available.

Antibody suppliers should always offer properly validated reagents, especially when providing phospho-specific antibodies. Ideally, this means showing a single band on a Western blot assay using total cell lysate rather than purified recombinant protein, and proof the antibody is phospho-specific. We at Novus Biologicals guarantee and provide proof of the purity and efficacy of our antibody reagents.