| Solution Information | help | |
| Enzyme: | Bcl-2-related protein A1 | |
| inhibitor: | BDBM53484 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Materials: Assay buffer: 25 mM Bis-Tris, 1mM TCEP, 0.005% Tween 20 Bfl-1: Produced at the Sanford-Burnham Medical Research Institute F-Bid: Produced at the Sanford-Burnham Medical Research Institute Tb x GST: Invitrogen (Catalogue #: PV3551) TCEP: Sigma (Catalogue #: 646547) Assay plate: Corning 1536 Well White Plate (Catalogue #: 3725) Protocol: I. Compound Addition: 1. Using LabCyte Echo, transfer 40 nL from a 10 mM Echo qualified plate containing test compounds into assay plate columns 5 - 48 (final concentration of test compounds is 100 microM, 1.0 % DMSO). Transfer 40 nL of DMSO to positive and negative control wells in columns 1 - 4. 2. Centrifuge plates at 1000 rpm for 1 min. 3. Seal the plates and leave them at RT. Note: Compounds are added to the plates before reagent addition II. Set up of Bfl-1 assay: 4. Prepare assay buffer 5. Bfl-1: Dilute Bfl-1 in assay buffer to make 2X intermediate solution at 10 nM 6. F-Bib/Tb x GST: Dilute F-Bid and Tb x GST in assay buffer to ma | |
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