Finding fragments is now routine, but how are people doing it? NMR of course has a venerable history, but X-ray crystallography provides higher resolution data, SPR is faster than either, and there are all sorts of other methods. To learn what’s state of the art (and to help me gather some data for an upcoming talk) please vote for what method(s) you’re using. Note that the vote is on the right side of the page, and you can vote for more than one method.
Affinity chromatography, capillary electrophoresis, or ultrafiltration
Computational screening
Functional screening (high concentration biochemical, FRET, etc.)
ITC (isothermal titration calorimetry)
MS (mass spectrometry)
NMR – ligand detected
NMR – protein detected
SPR (surface plasmon resonance)
Thermal shift assay
X-ray crystallography
Other – please specify in comments
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4 comments:
Anyone have any hints on how to increase S/N for thermal shift assays?
Better S/N for thermal shift assays (DSF):
Screen protein concentrations vs SYPRO Orange concentration.
Accept that some compounds just screw the assay.
Purer sample.
Apply Andrew Yeh's methods for background subtraction: http://www.ncbi.nlm.nih.gov/pubmed/16552147
Acquire a machine for differential static light scattering (DSLS).
Ordered by easiest to hardest/most cumbersome.
maybe that's obvious, but did you test different buffers?
and another rather easy thing: higher well volume (i.e. 40 instead of 20 uL)
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