Protein Identification

Overview

These services enable the identification of proteins in a gel band, gel spot or in solution. If you would like to identify a protein in a gel band or gel spot you should select our MSB-01 service. If you would like to identify a protein in solution you should select our MSB-02 service.

MSB-01 can be used to identify proteins in gel bands or gel spots. We can accept any type of gel matrix and most stains. If you are using a silver staining, please avoid the Bio-Rad Silver Stain Plus Kit. For the greatest sensitivity towards your target you should cut the band close to your target, the gel band dimensions will be ~5mm x 2mm. We can get an identification with as little as 1ng of protein on gel. There is a link at the bottom of the page that will let you view a Service Note that provides a useful sensitivity guide.

MSB-02 can be used to identify a protein in solution. We recommend submitting samples at a protein concentration of ≥1µg/µL in 10µL of buffer. This is just a guide, if you any concerns about the suitability of this service for the analysis of your samples please call or email. If you provide detailed buffer information with your samples it can be helpful. If not addressed during processing buffers containing detergents and non-volatile salts can interfere with the analysis.

Steps Involved

  • Sample preparation: In-gel digestion with trypsin using a ProGest (Digilab) robot. Solution digestion with trypsin is performed manually.
  • LC-MS/MS: Digested samples are analyzed with 0.5hr LC-MS/MS
  • Data analysis: Protein identification is performed with Mascot. Protein visualization and validation is performed with Scaffold.
  • Report generation: PDF/Excel/Scaffold

The turnaround time is 2-3 weeks.

Deliverables

  • Written Report containing details of your samples, our methods and a summary of the data.
  • An Excel workbook containing the detailed qualitative and quantitative protein identification data.
  • A Scaffold file to enable your own deeper data mining.

An example report is available upon request.

A service note with some data outlining some fundamentals of the service is available here.