Technical Reference #1959

1959.

Increased SOD1 association with chromatin; DNA damage; p53 activation; and apoptosis in a cellular model of SOD1-linked ALS Livea F. Barbosa; Fernanda M. Cerqueira; Antero F.A. Macedo; Camila C.M. Garcia; José Pedro F. Angeli; Robert I. Schumacher; Mari Cleide Sogayar; Ohara Augusto; Maria Teresa Carrì; Paolo Di Mascio ; Marisa H.G. Medeiros, Universidade de Sao Paulo, Biochimica et Biophysica Acta, 1802(1959), (2010)
Link To Paper

Keywords:
ALS SOD1 p53 Nucleus DNA damage

Materials & Methods:
Cells were seeded in 35-mm plates containing a glass coverslip (MatTek Ashland MA) fixed with paraformaldeyde (4%) for 15 min at room temperature and permeabilized with 0.1% Triton X-100. The samples were blocked for 1 h in 1% BSA in PBS and incubated for 2 h at 37 °C with sheep monoclonal anti-SOD1 antibody (Calbiochem Darmstadt Germany) and then washed in blocking buffer and incubated for 2 h with a goat anti-sheep antibody conjugated to Texas Red (Invitrogen Eugene OR). After rinsing in PBS the cellswere stained with 10 μg/mL DAPI (Invitrogen-Molecular Probes Eugene OR). The same protocol was performed for nuclear images except for the permeabilization step. All images were acquired with a Zeiss LSM 510 Confocal Microscope.

Microscopic Technique
Confocal microscopy

Cell Type(s)
SH-SY5Y human neuroblastoma cells