| Solution Information | help | |
| Enzyme: | Amyloid-beta A4 precursor protein-binding family A member 1 | |
| inhibitor: | BDBM54760 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | This assay was used as an alternate secondary screen to uHTS Homogeneous Time-Resolved Fluorescence Resonance Energy Transfer (HTRF) Assay AID 2073. Terbium HTRF in dose response Assay materials: 1) Glutathione S-transferase (GST)-fusion protein (GST-M15) from assay provider. 2) Fluorescein labelled N-type Ca2+ channel carboxyl-terminal peptide (fluorescein-NC peptide) corresponding to the carboxyl-terminal tail of CaV2.2 protein (Bio Peptide lot# 0610096426) 3) Anti-GST monoclonal antibody labeled with terbium Tb-anti-GST antibody (Invitrogen cat# PV3551). 4) Tween-20 (Sigma cat#9416) 5) Bovine Serum Albumin (Sigma cat# A7888) Working solutions: 1. Assay buffer: 25 mM HEPES, pH 7.2, 100mM NaCl,0.1% BSA, 0.005% Tween-20 2. Mint1-PDZ working solution (2X final concentration) contains 10 nM GST-MINT-PDZ and 20 nM Tb-anti-GST 3. Fluorescein-NC peptide- (2X final concentration) contained 0.5 nM fluorescein-NC peptide. 4. Unlabelled NC peptide-(2X final concentration) contained 0.5 n | |
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