MultiDrop Combi dispenser

Using MultiDrop Combi dispenser add 2 pL/well of 2x Assay Buffer to columns 1 and 2 (positive control wells). 

Using MultiDrop Combi Dispenser add 2 pL/well of 2x mlAP Solution to columns 3 through 48. 

With the Multidrop Combi dispenser and small metal tip cassette, dispense 3 pi Assay Buffer in columns 1-2 for Positive Control, and 3 pi diluted ENPPl in columns 3 8. With a different cassette, dispense 3 pi diluted ATP solution to all assay wells. Spin down the plate and seal with aluminum adhesive seal. Incubate for 110 min. 

Subsequently, a 1 pL per well substrate solution is dispensed to the assay plates in another Multidrop-Combi equipped with plate stackers. After 30-min incubation at room temperature, the operator manually moves the assay plates to a plate detector with stackers and executes the measurement of assay plates in a specific detection mode (fluorescence intensity in this case). The data file from the plate reader and the tracer file will be copied to a computer for data analysis after the experiment. This screening platform is very useful for laboratories that screen small compound collections, assay validation with the LOPAC collection, and follow-up screens for hit confirmation and lead optimization. 

Multidrop Combi Reagent Dispenser (Thermo Scientific, Waltham, MA, USA). 

Use Bravo liquid handler and one unused boxes of P30 tips to aspirate 19.5 pi from the blood intermediate plate and dispense 4.5 pi into quadruplicate occupied wells in the 1,536 well assay plate. Spin down plate and read on the PHERAstar plate reader at fixed time points, using 380 nm for pNPP substrate and 555 nm for PPMP substrate see Note 6). For PPMP substrate, use Multidrop Combi and small metal tip cassette to dispense 2 pi Color Developer solution and spin once more before reading. 

Dispense with Multidrop Combi and small metal tip cassette... 

Reagent dispenser, e.g.. Multidrop Combi (Thermo Scientiflc), Bravo Automated Liquid Handling Platform (Agilent Technologies). 

For automated HTS screening Multidrop Combi equipped with a small tube metal tip dispensing cassette for 0.5-50 pL volumes of kinase solution (Thermo Scientific). 

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