Milton RH, Abeti R, Averaimo S, DeBiasi S, Vitellaro L, Jiang L, Curmi PM, Breit SN, Duchen MR, Mazzanti M. CLIC1 function is required for beta-amyloid-induced generation of reactive oxygen species by microglia. J Neurosci. 2008 Nov 5;28(45):11488-99. PubMed Abstract

REAGENTS/MATERIAL:
Purity was assessed directly through immunofluorescent staining using antibodies against OX42 (microglia) and GFAP (astrocytes). CLIC1 localization was probed using antibodies to the protein and, more specifically, its extracellular N-terminus, after treatment with Aβ. The FLAG epitope is recognized by monoclonal anti-FLAG-m2 antibodies (Sigma-Aldrich). Cells were treated with Aβ 25–35 or vehicle for 1 h before fixation, permeabilization, and staining with antibodies to the CLIC1 protein and DAPI. Immunofluorescence of permeabilized BV2 cells was achieved using antibodies against CLIC1, both commercially sourced and laboratory generated.