The Thermofluor Screens JBScreen FUNDAMENT and JBScreen SPECIFIC allow identification of protein-stabilizing buffer conditions which is pivotal for protein purification, characterization and crystallization.
Undesired overlay of screening of interdependent variables is prevented by strictly categorizing stability screening of proteins into

1. FUNDAMENTAL factors that influence the whole protein molecule (pH and ionic strength)

2. SPECIFIC factors that affect energetically important hot spots on the protein (substrates and their analogs, cations, anions, ...)

The protein’s melting temperature (Tm) is used as reporter for protein stability and determined by a thermal shift assay (96-well plate format) monitoring the unfolding of a protein in a temperature-dependent manner. The higher the Tm, the higher is the thermostability of the protein in that specific environment.

JBScreen Thermofluor FUNDAMENT and SPECIFIC are provided in a deep-well block at 0.5 ml each, which allows a flexible application of this alternative approach.

References

• Ericsson et al. (2006) Thermofluor-based high-throughput stability optimization of proteins for structural studies. Anal. Biochem. 357(2):289.
• Reinhard et al. (2013) Optimization of protein buffer cocktails using Thermofluor. Acta Cryst. F 69:209.
• Niesen et al. (2007) The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Nat. Protoc. 2(9):2212.
• http://www.rcsb.org/pdb/home/home.do