The Durham pH Screen MD1-101


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The Durham pH Screen is a protein stability screen designed by Ehmke Pohl, Morten Grøftehauge and Emily Cardew at Durham University to be used with a protein stability assay, such as the thermal shift assay (also known as ThermoFluor®). The screen identifies how buffer chemistry and pH alters the thermal denaturation and therefore stability of a protein. In addition, buffer molecules can act as low affinity ligands and bind to specific pockets on a protein, this can be an undesirable confounding factor in fragment-based drug design projects. Therefore, a protein stability screen for the optimum buffer and pH for your sample is a vital first step in any structure determination project. The developers describe the importance of stability screening in this JoVE video article:

The Durham pH Screen includes 28 different buffers covering a broad pH range (4-11), and separates the effects of buffer chemistry and pH. We recommend it's use together with the Durham Salt and RUBICs screens to thoroughly understand the effect of protein environment on stability.

The Durham Screens are designed to work with NAMI* the free to download software for easy interpretation of thermal shift assay results.

Reference:Grøftehauge, MK, Hajizadeh, NR, Swann, MJ and Pohl, E. Protein-ligand interactions investigated by thermal shift assays (TSA) and dual polarization interferometry(DPI). Acta Cryst D71:36-44 (2015).

The Durham pH Screen is presented as 96 x 0.5 mL conditions in a deep-well block. They are preservative free and it is recommended that they are stored at 4°C, however they can be used at room temperature for screen set-up.

*NAMI can generate waterfall plots of thermal shift data at multiple concentrations or plots of melting temperature against concentration. These differentiate a sudden increase in protein stability indicating a specific interaction and steady increases in stability due to the changing protein environment. In addition, it produces a useful colour gradient plate diagram to make identifying particularly stabilising or destabilising conditions simple.

ThermoFluor is a registered trademark of Johnson and Johnson.




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