Protein purification

The CSB provides a variety of purification services that can be tailored to your research needs. These services can be provided in the form of personnel training or full contract services.

U-M Protein Purification and Crystallography Rates

If you are outside of U-M, please contact Jeanne Stuckey jass [at] umich.edu for rates.

Equipment

  • Multiple variable temperature shakers for both large- and small-scale fermentation
  • French press – for breaking E.coli cells
  • Sonicator with nano and macro tips for breaking E.coli cells
  • Microfluidizer - for breaking E.coli cells
  • Two Acta FPLCs equipped with a variety of ion exchange, affinity and both preparative and analytical sized gel filtration columns capable of purifying complexes up to 200,000 Da

Access is determined by Purification Protocol and Scale

To design a purification protocol, please contact Jeanne Stuckey at jass [at] umich.edu.
For small scale (<1 mg), one-step affinity purification, please contact Clay Brown at wclayb [at] umich.edu of the HTP lab.
For large scale (1 – 100 mg), single-step through multi-step purification using a known protocol, contact Jennifer Meagher at jmeagher [at] umich.edu.

Protein Purification FAQs

1. When should I call to schedule time on the FPLC?  Contact Jennifer Meagher at jmeagher [at] umich.edu or Priya Chinnaswarmy at kchinnas [at] umich.edu one to two weeks in advance.

2. How do I prevent my protein from aggregating during a concentration step? The V-style concentrators (Amicon Ultras) will cause concentration gradients to build up in your sample resulting in unwanted aggregation. To prevent this from occurring, spin for < 5 minutes, mix sample and repeat until the desired concentration is reached. If precipitation is still a problem, switch to the amicon centripreps, which concentrate at a slower rate without the development of a concentration gradient. If still plagued with precipitation problems, you may want to perform a protein stability profile on your sample to find a new buffer condition. Contact Jeanne Stuckey at jass [at] umich.edu or Priya Chinnaswamy at kchinnas [at] umich.edu for a quote.

 
 
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