Transcription Factor Sequestration by Polyglutamine Proteins - Protein Misfolding and Cellular Stress in Disease and Aging - page 210

4. TritonX-100.
5. Goat serum.
6. Mouse monoclonal antibody against V5 tag (R960-25;
Invitrogen).
7. Anti-mouse IgG conjugated with Alexa 546 (Molecular
Probes).
8. 4¢-6-Diamidino-2-phenylindole (DAPI).
9. VECTASHIELD Mounting Medium (Vector Laboratories).
10. TBST (20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.05%
Tween20).
11. Fluorescence microscope or confocal laser microscope.

1. Aminopropylsilane (APS) coated slide glass.
2. Phosphate buffered saline.
3. 4% PFA in PBS.
4. Bovine serum albumine (BSA).
5. Freezing microtome.
6. Mouse monoclonal anti-ubiquitin antibody (MAB1510;
Chemicon International) or rabbit polyclonal anti-ubiquitin
antibody (Z0458; DAKO).
7. Antibody against transcriptional factor.
8. Secondary antibodies conjugated with Alexa 488 or Alexa
546 (Molecular Probes).
9. 4¢-6-Diamidino-2-phenylindole (DAPI).
10. VECTASHIELD Mounting Medium (Vector Laboratories).
11. TBST (20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.05%
Tween20).

2.2. Materials
for Experiments Using
HD Model Mouse Brain

2.2.1. Materials for
Immunofluorescence
Microscopy

1. Sense and antisense oligonucleotides containing transcrip-
tional factor binding site.
2. g[ 32 P]-ATP (6,000 Ci/nmol).
3. T4 DNA kinase and kinase buffer.
4. Sepharose G-50 spin column (Amersham).
5. Lysis buffer (20 mM Hepes, pH 7.9, 25% glycerol, 150 mM
NaCl, 1.5 mM MgCl 2 , and Complete protease inhibitor
(Roche)).
6. Glass homogenizer.
7. Buffer C (20 mM Hepes, pH 7.9, 25% glycerol, 420 mM
NaCl, 1.5 mM MgCl 2 , and Complete protease inhibitor).
8. 2× binding buffer (20 mM Tris-HCl, pH 7.5, 100 mM NaCl,
2 mM EDTA, 10% glycerol).

2.2.2. Materials for
Electrophoretic Mobility
Shift Assay