A high-throughput MALDI MS assay for multiplexed read-out of histone demethylase inhibition
| Type: | Application |
Poster |
Expression Proteomics |
| Number: | Technology |
ASMS 2010, ThP24 553 |
MALDI-TOF |
| Year | Products |
| Author | |
Hamzah N Freeman1, Melanie Leveridge1, Sue Hutchinson1, Andy West1, Anja Resemann2, Detlev Suckau2, Klaus Schneider1 1Molecular Discovery Research, GlaxoSmithKline, New Frontiers Science Park, Harlow and Medicine Research Centre, Stevenage UK and 2Bruker Daltonics, Bremen, Germany |
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| Reference | |
Abstract |
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•A robust and high-throughput MALDI-TOF MS assay has been developed for defining histone methylation status suitable for screening of inhibitors of histone demethylase function by direct detection of substrate and product. •Automated protocol established for sample preparation from 384-well plate onto matrix pre-coated 384 spot target plates. • Assay Z prime range of 0.6 - 0.8 for a typical analysis. •Data collection time 2 second per spot using a fixed laser power at 1 kHz firing rate (smartbeam II laser) on the new autoflex speed MALDI-TOF allows rapid read-out of a 384-well target in 13 minute. •Multiplexing proof-of-concept for the assay has been shown for the duplex case. Multiplexing will lead to further increased throughput of read-out and reduced cost-per-data point. •The assay method is capable of high-throughput read-out of compound effect on demethylase function for driving lead optimisation in drug discovery. |
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