| Solution Information | help | |
| Enzyme: | DNA repair protein RAD51 homolog 1 | |
| inhibitor: | BDBM43226 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Materials Human Rad 51 protein, labeled ss & ds DNA were provided by the assay provider. The fluorescence assay was carried out in 384-well black, low-volume plates from Corning (Cat # 3676). All buffer salts were from Sigma. Assay hRad 51 protein was incubated with ss DNA to allow filament formation (37 deg C for 15 min). Compounds (8.5 uM final concentration) were pre-incubated with the protein for 30 min. followed by addition of ds DNA to initiate strand exchange. The reaction was allowed to proceed at room temperature for 15 min and then plates were read on Envision plate reader. Dose response protocol 1.Serial dilute single compounds at in DMSO (16 two-fold dilutions from 2.5 mM to 75 nM) 2.Fill 384 well plate with 4 uL of water (nuclease-free) using Multidrop 3.Pin Tool compound into the plates using 384 pin-tool 4. Add 4 ul reaction mix (containing hRad 51 and ss DNA with buffers) 5.Preincubate compound with the reaction mix for 30 min at room temperature 6. Add 2 ul ds D | |
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