MST screen for RNA binding inhibitors

RBD binding to Inhibitor conditions, Julia Welte

How to set up the inhibitor screen with the picoMST or NT115

which machine

does not really matter - which was used for establishement. what is the occupancy.

inhibitors

Inhibitors to test: RBD inhibitors 1st round - PharmAI

Inhibitors have to be plated by 2bind

first round: final concentration in reaction 100 uM

depending on the number of positives - reduce concentrations

RNA to use

full-length NP: the double stranded DNA shows a clear cooperative binding signal - protein 2uM

signal about 30 fluorescence units

RBD-domain: ????

full-length or RBD?

the screen was against the RBD - better to use this. and then the positive hits as well against the full-length.

protein batch to use

date of production

buffer condition

.....

MST screen setup

capillary 1 to 16

1 DMSO+RNA / 2 DMSO+RNA+Protein

3-14 DMSO+Inhibitor+RNA+Protein in singlett (12 compounds per round)

15 DMSO+RNA / 16 DMSO+RNA+Protein

about 22 runs

buffer conditions to use

xxxx

RNA-protein concentration to use

xxxxx