Caspases,Paracaspases, and Metacaspases Methods and Protocols

148 Incubate with secondary antibody (e.g., goat anti-rabbit HRP- conjugated IgG; Santa Cruz cat# sc-2004) diluted 1:10,000 in ...

149 ischemic brain [ 24 ], a controlled cortical impact (CCI) model [ 25 ], and conjunctival epithelia in B6 mice [ 26 ]. The av ...

150 Incubate the sections with caspase antibody at appropriate dilution (at least 1:200) in antibody dilution buffer in a humid ...

151 At present many target substrates of caspases have been identifi ed in vitro and in vivo [ 27 ]. Proteins cleaved by caspase ...

152 should be examined. The time of incubation and the amount of substrate concentration should be used for the assay within the ...

153 Photobleaching is a common problem during immunofl uores- cence studies. This can be controlled by reducing the time- span ...

154 against caspase-induced neo-epitopes. Methods 44:255–261 Ohsawa S, Hamada S, Yoshida H et al (2008) Caspase-mediated change ...

155 Chapter 10 Measurement of Caspase Activation in Mammalian Cell Cultures Magnus Olsson and Boris Zhivotovsky Abstract The maj ...

156 sequential activations form the foundation for apoptotic cascades. The catalytic activity of an effector caspase is enhanced ...

157 Several theoretical models have been postulated aiming to clarify the molecular mechanism by which initiator caspases become ...

158 cell death process while others, since they happen to contain a caspase target motif in their sequence, may be cleaved as by ...

159 molecule (M30) to which monoclonal antibodies have been raised [ 11 ]. Unconjugated primary antibody targeting the neoepitop ...

160 Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-fl uoromethylk- etone) is a pan-caspase inhibitor that irreversibly binds to t ...

161 Electrode (running) buffer (10×): Dissolve 30 g of Tris base (247 mM), 144 g glycine (1.92 M), and 10 g sodium dodecyl sulf ...

162 Glass Coverslips: 12 mm circles (fi t in 12 or 6-well culture plates), Thickness No. 1. Coverslips should be cleaned with e ...

163 Anti-Cytokeratin 18 M30 CytoDEATH Monoclonal Antibody, FITC Conjugated ( Roche Applied Science ). Antibody dilution buffer: ...

164 Caspase inhibitor stock solutions: Prepare stock solutions of each specifi c caspase inhibitor (Z-VAD-FMK, Z-XXXD-FMK, or Z- ...

165 Pour the separating gel in between the glass plates. Leave suffi cient space for the stacking gel to be added later (the le ...

166 Place the cassette into electrode assembly and fi ll the tank with cold transfer buffer. Connect the electrophoresis unit t ...

167 All incubations should be carried out at room temperature unless otherwise noted, in a humid covered dish or plate to preven ...