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The RUBIC thermofluor screens aimed at helping determine protein stability, and ideal purification & storage conditions. They suit a wide range of proteins and can also be used to optimize protein samples for NMR experiments.

 

 
RUBIC
Developed in 2010 by Stephane Boivin and Rob Meijers at the Sample Preparation and Characterisation (SPC) facility at EMBL Hamburg, the "ready-to-use" Thermofluor screens readily became highly-sought after by the facility users.
  

 

The RUBIC Buffer screen is designed to screen for global parameters, and the RUBIC Additive screen – designed to search for protein-specific additives. The Buffer screen contains diverse buffers commonly used in structural biology experiments, and is designed in such a way, that it is possible to discern global stability trends according to pH, salt concentration, buffer type and concentration.

The RUBIC Additive screen contains small molecules that can affect the folding, aggregation state and solubility of the protein construct and also includes small molecules that specifically bind and stabilise proteins.

The Buffer and Additive screens can be used in synergy in a two-step approach: first screening for global parameters, then searching for protein-specific additives. These screens have been tested on more than 300 different protein constructs at the Sample Preparation and Characterization (SPC) facility at EMBL Hamburg and refined to suit a wide range of proteins.

Screen composition available at Molecular Dimensions website:

  MD

 

Sample requirement:

~210ul of protein in low ionic sample buffer free of stabilizing reagent. Initial protein at 10 - 20uM (35kDa) is normally sufficient to visualize a melting curve with a good signal-to-noise ratio. Lower concentration can be used with larger protein, while smaller protein may requires more concentrated protein. Sample buffer should contains reagent to stabilize protein, we recommend not to exceed NaCl (<200uM), glycerol (<10%), reducing reagent (<5mM). Assay is not compatible with most of the detergents.

"The RUBIC Screens are available at the SPC as prefilled ready-to-use microplate".