How to detect protein ubiquitination

Ubiquitination is a post-translational modification of proteins, referring to the process where the small polypeptide ubiquitin (Ub) is covalently attached to substrate or target proteins under the action of related enzymes. Ubiquitination primarily mediates protein degradation, but with in-depth research on protein ubiquitination, it has been found that ubiquitination can also participate in regulating other vital activities, such as the cell cycle, transcription activation, gene silencing, and immune response.

So how can we detect whether a target protein has undergone ubiquitination?

ProteinUbiquitination Detectionincludes whether sample proteins have undergone ubiquitination, what type of ubiquitination has occurred, and how many proteins have been ubiquitinated. Currently, the main methods for detecting protein ubiquitination are based on specific antibodies and mass spectrometry.

Methods based on specific antibodies include chromatin immunoprecipitation and Western blotting. Their basic principles are similar: extracting, purifying, and separating sample proteins, then combining them specifically with antibodies, followed by color development and imaging to analyze the ubiquitination status of proteins.

Since the degree of protein ubiquitination is low under normal physiological conditions, before mass spectrometry identification, sample proteins are usually digested, enriched using specific recognition antibodies for ubiquitinated proteins, followed by mass spectrometry analysis. Finally, mass spectrometry data are compared with databases and analyzed using bioinformatics for qualitative and quantitative identification of protein ubiquitination.

Biotech company Biotage uses Thermo Fisher's Q Exactive HF mass spectrometry platform combined with Nano-LC to launchProtein Ubiquitination Detectionservice package. You only need to tell us your experimental objectives and send us your samples; we will handle all subsequent matters of the project, including protein extraction, proteolysis, ubiquitinated peptide enrichment, peptide separation, mass spectrometry analysis, raw mass spectrometry data analysis, and bioinformatics analysis.

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