Post-translational Modifications (PTMs)

 

Disulphide Bridge Analysis

This technique is used to characterise disulphide folding pattern, i.e., an analysis of which Cys residues are linked.

Scope of work:

  • Confirmation of Protein identification by MS. We will provide unambiguous identification using our standard MS/MS sequencing service.
  • Identification of location of disulphide bonds. Full protein characterisation will be performed using our high-end LC-MS/MS and/or MALDI-TOF/TOF instruments. The analysis will look for di-sulphide bonded peptide fragments following enzyme digestion of the protein.

For analysis we require the full theoretical protein sequence, and postulated S-S bridging patterns.

Request Form: Disulphide Bridge Analysis

 

Phosphorylation Detection

Identification of phosphorylation sites on protein and peptide samples is achieved through precursor ion and neutral loss scans that allow detection of the loss of the phosphate group. From this data, phosphorylation sites can be mapped to detect this important post-translational modification of proteins.

For phosphorylation detection we use a LC-MS/MS system (4000Q-Trap [Sciex]). Our approach is first to perform protein identification by tandem MS (as per our standard service). This enables confirmation of protein identity prior to detection of the phosphorylated peptides.

Request Form: Phosphorylation Detection

 

Frequently Asked Questions

How much protein do I need?

We recommend protein bands or spots are of sufficient concentration to be detected by Coomassie G250 staining. Two bands are required to enable the initial protein identification, although less material can be used for that process. Larger amounts of material increase the probability of achieving a successful result.

 

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