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Protein Production in E. coli with Isotope Labelling for NMR, CERM/CIRMMP, Florence, Italy

The isotope labelling platform at CERM/CIRMMP is devoted to the large scale production of proteins uniformly enriched in stable isotopes (15N, 13C and 2H) for biomolecular NMR spectroscopy studies. The platform is offering protein expression in bacterial cultures grown on chemically defined minimal media, as well as advanced protein purification approaches with the unique possibility of running them in anaerobicity. Users have access to the facility for large scale expression of labelled proteins in E. coli, under the supervision of a qualified platform engineer. Dedicated bench, optimised protocols and adequate isotopically labelled materials are available to users.

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User Guide

CERM/CIRMMP isotope labelling platform combines a large set of expression vectors for Escherichia coli and protocols, which enable a broad screening of soluble protein expression in labelled media through the use of multiple fusion proteins and various growth conditions.

CERM/CIRMMP isotope labelling expression platform offers two modes of access

  1. The platform provides the expression vectors to the external users who perform cloning at their home institutions and come to CERM/CIRMMP to perform the expression assays in minimal media
  2. The external users perform cloning and expression assays in minimal media at the CERM/CIRMMP platform

Support includes training and supervision, as well as help with data analysis which is especially crucial for complex systems. The facility has a long-term expertise in producing metallo-proteins.

Cost For Users

Approved Instruct Users: Access costs (Consumables, travel and accommodation) can be partially or fully covered by Instruct, depending on the general costs.

Academics: The cost of isotopic materials and consumables used for the production of proteins will be refunded by the users to the CERM/CIRMMP platform.

Private companies: quotation under request.

How to make a request

A request form should be sent to the administrators of the platform by email. Following receipt of the form, we will study the feasibility of your request within one month. If the request is accepted, we will contact you in order to propose a date for the access to the platform. Alternatively if a request cannot be satisfied, external users can have access to our protocols in order to express the protein in their lab.