| Solution Information | help | |
| Enzyme: | Huntingtin | |
| inhibitor: | BDBM37572 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Material: For the culture and the screen - Established cell lines from Dr. Ai Yamamoto: exon1htt(103Q)mCFP (frozen stock in cell bank) - Trypsin/EDTA (GibcoBRL Invitrogen, #25300-112) - DMEM (GibcoBRL Invitrogen, #11960-077) - Sterile PBS (Tissue Culture) - Hygromycin B (BD Biosciences, 50 mg/ml) - Genetycin (G418) (Invitrogen) - Compound library - Positive control: Doxycycline (Sigma, #D9891-1g) [100 ng/ml final concentration] - Paraformaldehyde (EMS, #15710 10x10 ml) - Packard Plates (Perkin Elmer, #6005182) containing cells ready to be processed For the Detection: - Phosphate Buffered Saline (PBS), tablets, w/o Ca2+ and Mg2+ (MP Biochemicals, #2810305) - Hoechst 33342 (Molecular Probe Invitrogen, #H3570, 10 mg/ml solution in water) Working solution: - Fixation Solution: 4% Paraformaldehyde in PBS - PBS - Hoechst 33342 [1:10,000 (1 microM) in PBS]: 50 microl per well X 100 X number of plate+ an additional 10%) Procedure: Cell Seeding - Cells are cultured in DMEM supplemented with | |
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